CN105246915A - HGF antibody and composition containing same - Google Patents

Abstract

The present invention is directed to antibodies and fragments thereof having binding specificity for HGF. Another embodiment of this invention relates to the antibodies described herein, and binding fragments thereof, comprising the sequences of the VH, VL and CDR polypeptides described herein, and the polynucleotides encoding them. The invention also contemplates conjugates of anti-HGF antibodies and binding fragments thereof conjugated to one or more functional or detectable moieties. The invention also contemplates methods of making said anti-HGF antibodies and binding fragments thereof. Embodiments of the invention also pertain to the use of anti-HGF antibodies, and binding fragments thereof, for the diagnosis, assessment and treatment of diseases and disorders associated with HGF.

Description

HGF antibody and composition containing same

This application claims priority to US Provisional Serial No. 61/782,868 and US Provisional Serial No. 61/781,643, both filed March 14, 2013. The contents of these applications are incorporated herein by reference in their entirety.

This application includes as part of its disclosure the Biological Sequence Listing concurrently submitted via EFS-Web. The biological sequence listing is contained in a file named "43257o4013.txt", which was created on March 12, 2014 and is 652,047 bytes in size, and is incorporated by reference in its entirety In this article.

Background of the invention

field of invention

The present invention relates to antibodies and fragments thereof, preferably high affinity or avidity antibodies with binding specificity for hepatocyte growth factor (hereinafter referred to as "HGF"). The present invention also relates to methods of screening for diseases and disorders associated with HGF, and methods of preventing or treating diseases and disorders associated with HGF by administering said antibodies or fragments thereof.

Description of related technologies

Hepatocyte growth factor (HGF), also known as scatter factor (SF), is produced as a single-chain inert precursor that is cleaved by serine proteases into two chains linked by a disulfide bond. (Abounader, R. et al., Neuro-Oncology, 7:436-451 (2005)). The gene encoding HGF is located on chromosome 7q21.1. The biologically active form of HGF is a heterodimer composed of a 69-kDa alpha chain and a 34-kDa beta chain. The alpha chain contains an N-terminal hairpin domain and four kringle domains, while the beta chain contains a serine protease-like domain that is not enzymatically active. References ibid.

Human hepatocyte growth factor (HGF) is a multifunctional heterodimeric polypeptide produced by mesenchymal cells. HGF has been shown to stimulate angiogenesis, morphogenesis, and motogenesis, as well as growth and dispersion of various cell types (Bussolino et al., J. Cell. Biol. 119:629, 1992; Zarnegar and Michalopoulos, J. Cell .Biol.129:1177,1995; Matsumoto et al., Ciba.Found.Symp.212:198,1997; Birchmeier and Gherardi, Trends Cell.Biol.8:404,1998; Xin et al., Am.J.pathol.158 :1111,2001). The pleiotropic activity of HGF is mediated by its receptor, a transmembrane tyrosine kinase encoded by the proto-oncogene c-met. In addition to regulating various normal cellular functions, HGF and its receptor c-met have been shown to be involved in tumor initiation, invasion and metastasis (Jeffers et al., J. Mol. Med. 74:505, 1996; Comoglio and Trusolino, J. Clin. Invest. 109:857, 2002). HGF/c-met is coexpressed, often overexpressed, on a variety of human solid tumors, including those derived from lung, colon, rectum, stomach, kidney, ovary, skin, multiple myeloma, and thyroid tissue ( People such as Prat, Int.J.Cancer49:323,1991; People such as Chan, Oncogene2:593,1988; People such as Weidner, Am.J.Respir.CellMol.Biol, 8:229,1993; People such as Derksen, Blood99: 1405, 2002). HGF acts as an autocrine (Rong et al., Proc. Natl. Acad. Sci. USA 91:4731, 1994; Koochekpour et al., Cancer Res. 57:5391, 1997) and paracrine growth factor (Weidner et al., Am. J. Respir. Cell Mol. Biol. 8:229, 1993) and anti-apoptotic regulators (Gao et al., J. Biol. Chem. 276:47257, 2001).

HGF is a 102kDa protein whose sequence and structure are similar to plasminogen and other thrombin (Nakamura et al., Nature 342:440, 1989; Weidner et al., Am.J.Respir.Cell.Mol.Biol.8 :229, 1993, each of which is incorporated herein by reference). Human HGF is synthesized as a 728 amino acid precursor (preproHGF) that is cleaved intracellularly to an inert single-chain form (proHGF) (Nakamura et al., Nature 342:440, 1989: Rosen et al., J. Cell. Biol .127:1783, 1994). After extracellular secretion, proHGF is cleaved to produce a biologically active disulfide-linked heterodimeric molecule composed of α subunit and β subunit (Nakamura et al., Nature 342:440, 1989; Naldini et al., EMBOJ.11 :4825,1992). The alpha subunit contains 440 residues (69 kDa glycosylation) and consists of an N-terminal hairpin domain and four kringle domains. The β subunit contains 234 residues (34 kDa) and has a serine protease-like domain that lacks proteolytic activity. HGF cleavage is required for receptor activation, but not for receptor binding (Hartmann et al., Proc. Natl. Acad. Sci. USA 89:11574, 1992; Lokker et al., J. Biol. Chem. 288:17145, 1992). HGF contains 4 putative N-glycosylation sites, 1 in the alpha subunit and 3 in the beta subunit. HGF has 2 unique cell-specific binding sites: a high-affinity (Kd=2×10 ^-10 M) binding site for c-met receptor and a low-affinity (Kd=10 M) binding site for heparan sulfate proteoglycan (HSPG). ^-9 M) binding sites, which are present on the cell surface and on the extracellular matrix (Naldinl et al., Oncogene 6:501, 1991; Bardelii et al., J.Biotechnol.37:109, 1994; Sakata et al., J.Biol. Chem., 272:9457, 1997).

c-met is a member of the class IV protein tyrosine kinase receptor family. The full-length c-met gene was cloned and identified as a c-met proto-oncogene (Cooper et al., Nature 311:29, 1984; Park et al., Proc. Natl. Acad. Sci. USA 84:6379, 1987). The c-met receptor was originally synthesized as a partially glycosylated single-chain precursor p170 _(MET) (Park et al., Proc. Natl. Acad. Sci. USA84:6379, 1987; Giordano et al., Nature 339:155, 1989 ; Giordano et al., Oncogene 4:1383, 1989; Bardelli et al., J. Biotechnol. 37:109, 1994). After further glycosylation, the protein is proteolytically cleaved into a heterodimeric 190 kDa mature protein (1385 amino acids) consisting of a 50 kDa alpha subunit (residues 1-307) and a 145 kDa beta subunit. The cytoplasmic tyrosine kinase domain of the β subunit is involved in signal transduction.

Several different approaches have been investigated to obtain HGF inhibitors or HGF antagonists. Such inhibitors include truncated HGF proteins such as NK1 (N-terminal domain plus kringle domain 1: Lokker et al., J. Biol. Chem. 268:17145, 1993); NK2 (N-terminal domain plus kringle domain 1 and 2: Chan et al., Science 254:1382, 1991); and NK4 (N-terminal domain plus four kringle domains), which were shown to partially inhibit primary growth and metastasis of murine lung tumor LLC in a nude mouse model (Kuba et al., Cancer Res. 60:6737, 2000).

As another approach, Dodge (M.S. Thesis, San Francisco State University, 1998) generated antagonist anti-c-met monoclonal antibodies (mAbs). One mAb, 5D5, exhibited strong antagonistic activity in ELISA, but induced a proliferative response in c-met-expressing BAF-3 cells, possibly due to dimerization of membrane receptors. For this reason, a single domain form of the anti-c-metmAb 5D5 has been developed as an antagonist (Nguyen et al., Cancer Gene Ther, 10:840, 2003).

Cao et al., Proc.Natl.Acad.Sci.USA98:7443,2001 reported that in a xenograft nude mouse model, administration of a mixture of three anti-HGF mAbs (selected based on their ability to inhibit HGF dispersal activity in vitro) was able to inhibit human Tumor growth.

Recently, neutralizing (inhibitory) anti-HGF mAbs have been reported, including L2G7 (Kim et al., Clin Cancer Res 12:1292, 2006, WO2005/107800 and USP 7,220,410), HuL2G7 (WO2007/115049), human described in WO2005/17107. mAbs and HGF binding proteins as described in WO2007/143090 or WO2007/143098. It has also been reported that anti-HGFmAbL2G7, when administered systemically, strongly inhibits the growth of orthotopic (intracranial) glioma xenografts or even induces their regression and prolongs animal survival (Kim et al., supra and WO2006/ 130773).

As disclosed herein, HGF promotes the growth and/or dispersion of various cell types and has been shown to be involved in promoting angiogenesis, inhibiting cell growth, and transitioning from mesenchymal to epithelial phenotypes. Furthermore, both HGF and c-met are expressed in various human tumors, and their expression levels are sometimes associated with poor prognosis. Furthermore, HGF is thought to play a role in the development of a variety of diseases and conditions including, but not limited to, the development and metastasis of many cancers, and the development of macular degeneration. Due to the perceived involvement of HGF in various diseases and conditions, there remains a need in the art for HGF antagonists, compositions thereof and methods useful in the prevention or treatment of HGF-related diseases, as well as for identifying patients with HGF-related diseases or conditions screening methods for patients. Particularly preferred are anti-HGF antagonists and compositions thereof that effectively inhibit at least one HGF-related biological activity when administered to a patient with minimal or no adverse effects. The present invention achieves this goal.

Summary of the invention

The present invention relates to specific antibodies and fragments thereof having binding specificity for HGF, in particular antibodies having desired epitope specificity, high affinity or avidity and/or functional properties. One embodiment of the invention encompasses chimeric or humanized antibodies and fragments thereof capable of binding to HGF and/or HGF-HGFR complexes. Another embodiment of the invention relates to the antibodies described herein comprising the sequences of the _VH , _VL and CDR polypeptides described herein, and polynucleotides encoding them. In more particular embodiments of the invention, these antibodies will have a binding affinity (Kd) of less than 500 picomolar and/or a dissociation rate constant (K _off ) value of less than or equal to 10 ⁻⁴ S ⁻¹ .

More specifically, the invention provides rabbit antibodies with specificity for HGF and humanized and chimeric antibodies derived therefrom as well as antibody fragments with specificity for HGF including, for example, Fab', F(ab' ) ₂ , Fv, scFv fragments, SMIPs (Small Molecule Immunopharmaceuticals), Camelids, Nanobodies, monovalent antibodies (such as MetMab-like antibodies) and IgNARs, which can be used in therapy and diagnostics.

In addition, the invention provides nucleic acids encoding and promoting the expression of anti-HGF antibodies (i.e., rabbit antibodies) and antibody fragments of the invention, and modified forms thereof, including, for example, human derived therefrom, and host cells containing said nucleic acids. Fab and chimeric antibodies as well as antibody fragments including, for example, Fab', F(ab') ₂ , Fv, scFv fragments, SMIPs (Small Molecule Immunopharmaceuticals), Camelid antibodies, Nanobodies, monovalent antibodies (such as MetMab-like antibodies), and IgNAR.

The present invention also relates to expression systems for producing the anti-HGF antibodies of the present invention, including yeast, fungi, mammals and other cells suitable for producing antibodies and antibody fragments.

Furthermore, the present invention relates to novel antibodies and antibody fragments that specifically bind to human HGF that compete for and/or specifically bind to the same or overlapping epitopes on HGF as any of the anti-HGF antibodies and antibody fragments exemplified herein.

In addition, the present invention provides anti-HGF antibodies and antibody fragments, which partially or completely neutralize HGF, and which partially or completely inhibit one or more biological activities of HGF, such as HGF causing fibrosis or dispersion of cells, proliferation, angiogenesis or chemotaxis capacity.

The invention also relates to the in vivo use of the anti-HGF antibodies and antibody fragments of the invention, alone or in combination with other active agents or drugs, for blocking, inhibiting or neutralizing HGF or at least one activity of HGF and/or for inhibiting or Block HGF/HGF-R (c-met) interaction or inhibit c-met activation.

The present invention also particularly relates to the in vivo use of the anti-HGF antibodies and antibody fragments of the present invention alone or in combination with other active agents or drugs.

The present invention also particularly relates to the use of the anti-HGF antibodies or fragments thereof as described herein for ameliorating or alleviating the symptoms of, or treating or preventing, the following non-limiting list of proliferative, non-proliferative diseases and disorders A non-limiting list, such as cancers, including ovarian cancer, breast cancer, lung cancer (small cell or non-small cell), colon and colorectal cancer, prostate cancer, pancreatic cancer, kidney cancer, gastric cancer, liver cancer, bladder cancer, thyroid cancer, Endometrial cancer, head and neck tumors, melanoma, sarcoma, leukemia; lymphoma; and brain tumors (eg, glioblastoma) in children or adults; macular degeneration; sdisease); and malaria infection. In a preferred embodiment, the disease is selected from cancer or macular degeneration.

The invention also relates to agents for the treatment and/or prophylactic treatment of various diseases such as cancer, tumors, cell proliferative disorders, immune (such as autoimmune) disorders and/or angiogenesis-related disorders. In another embodiment, the present invention provides the use of a nucleic acid of the present invention in the manufacture of a medicament for the treatment and/or prophylaxis of diseases such as cancer, tumors, cell proliferative disorders, immune (such as autoimmune ) disorders and/or angiogenesis-related disorders.

The present invention also relates to the use of the expression vector of the present invention in the preparation of a medicament for the treatment and/or prophylaxis of diseases such as cancer, tumors, cell proliferative disorders, immune (such as autoimmune) disorders and/or Angiogenesis-related disorders. In another embodiment of the present invention, the present invention provides the use of the host cell of the present invention in the preparation of a medicament for the treatment and/or prophylaxis of diseases such as cancer, tumors, cell proliferative disorders, immune ( such as autoimmune) disorders and/or angiogenesis-related disorders. In another embodiment of the invention, the invention provides the use of the preparation of the invention for the manufacture of a medicament for the treatment and/or prophylaxis of diseases such as cancer, tumors, cell proliferative disorders, immune (such as autoimmune) disorders and/or angiogenesis-related disorders. The invention also relates to the use of the kit of the invention for the preparation of a medicament for the treatment and/or prophylaxis of diseases such as cancer, tumors, cell proliferative disorders, immune (such as autoimmune) disorders and/or Angiogenesis-related disorders.

The present invention is also particularly concerned with modulating conditions associated with dysregulation of the HGF/c-met signaling axis and thereby modulating at least one of cell proliferation, invasion, metastasis and angiogenesis.

The methods of the invention can be used to affect any pathological condition associated with dysregulation of HGF/c-met signaling, including chronic and acute conditions or diseases, including those pathological conditions that predispose mammals to the condition in question. Non-limiting examples of conditions to be treated herein include malignant and benign tumors; non-leukemic and lymphoid malignancies; neuronal, glial, astrocyte, hypothalamic and other glandular, macrophage, epithelial Cellular, stromal, and blastocoel disorders; and inflammatory, immune, and other angiogenesis-related disorders. Carcinoma, lymphoma, blastoma, sarcoma and leukemia. More specific examples of such cancers include squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, peritoneal carcinoma, hepatocellular carcinoma, gastrointestinal cancer, pancreatic cancer, colloid Blastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney cancer, liver cancer, prostate cancer, Vulvar cancer, thyroid cancer, liver carcinoma, and various types of head and neck cancer; disorders involving dysregulation of angiogenesis, including non-neoplastic and neoplastic conditions, such as the cancers and non-neoplastic conditions described herein, including but not limited to ) undesired or abnormal hypertrophy, arthritis, rheumatoid arthritis (RA), psoriasis, psoriatic plaques, sarcoidosis, atherosclerosis, atherosclerotic plaques, diabetic and other hyperplasia Retinopathy, including retinopathy of prematurity, retrolentic fibroplasia, neovascular glaucoma, age-related macular degeneration, diabetic macular edema, corneal neovascularization, corneal graft neovascularization, corneal graft rejection, retinal/choroidal Neovascularization, neovascularization in the corner of the eye (iridosis), ocular neovascular disease, vascular restenosis, arteriovenous malformation (AVM), meningioma, hemangioma, angiofibroma, thyroid hyperplasia (including Graves' Grave's disease), corneal and other tissue transplantation, chronic inflammation, pulmonary inflammation, acute lung injury/ARDS, sepsis, essential pulmonary hypertension, malignant pulmonary effusion, cerebral edema (e.g. associated with acute stroke/atresia) head injury/trauma-related), synovial inflammation, pannus formation in RA, myositis ossificans, hypertrophic bone formation, osteoarthritis (OA), refractory ascites, polycystic ovarian disease, uterine Endometriosis, 3rdspacingoffluiddisease (pancreatitis, compartment syndrome, burns, enteropathy), uterine fibroids, prematurity, chronic inflammation such as IBD (Crohn's disease) and ulcerative colitis), renal allograft rejection, inflammatory bowel disease, nephrotic syndrome, unwanted or abnormal mass growth of tissue (not cancer), hemophagocytic joints, hypertrophic scarring, inhibited hair growth, Osler-Weber syndrome, pyogenic granuloma, retrolentic fibroplasia, scleroderma, trachoma, vascular adhesion, synovitis, dermatitis, preeclampsia, ascites, pericardial effusion ( Such as pericardial effusion associated with pericarditis) and pleural effusion.

In certain embodiments, specific binding agents to HGF are used with one or more specific therapeutic agents to treat various cancers. In certain embodiments, a specific binding agent to HGF is used with one or more specific therapeutic agents to treat or prevent malaria. In certain embodiments, a specific binding agent to HGF is used with one or more specific therapeutic agents to treat or prevent proliferative diabetic retinopathy. In certain embodiments, two, three, or more agents may be administered, given the condition and degree of treatment desired. In certain embodiments, the agents may be provided together by inclusion in the same formulation. In certain embodiments, the agent and the specific binding agent to HGF may be provided together by inclusion in the same formulation. In certain embodiments, the agents may be formulated separately and provided together by inclusion in a therapeutic kit. In certain embodiments, the agent and the specific binding agent to HGF may be formulated separately and provided together by inclusion in a therapeutic kit. In certain embodiments, the agents may be provided separately.

Conjugates of anti-HGF antibodies and binding fragments thereof conjugated to one or more functional or detectable moieties are also encompassed by the invention. The invention also encompasses methods of making said chimeric or humanized anti-HGF or anti-HGF/HGFR complex antibodies and binding fragments thereof. In one embodiment, binding fragments include, but are not limited to, Fab, Fab', F(ab') ₂ , Fv, scFv fragments, SMIPs (Small Molecule Immunopharmaceuticals), Camelids, Nanobodies, and IgNAR.

Embodiments of the present invention relate to the use of anti-HGF antibodies for the diagnosis, evaluation and treatment of diseases and disorders associated with HGF or its abnormal expression. The present invention also encompasses the use of fragments of anti-HGF antibodies for the diagnosis, evaluation and treatment of diseases and disorders associated with HGF or its aberrant expression. Other embodiments of the invention relate to the production of anti-HGF antibodies in recombinant host cells, preferably diploid yeast, such as diploid Pichia and other yeast strains.

Brief description of the drawings

Figure 1 provides human HGF ELISA binding data for antibody Abl obtained following the protocol in Example 8 below.

Figure 2 provides the human HGF ELISA binding data for antibody Ab2 obtained following the protocol in Example 8 below.

Figure 3 provides human HGF ELISA binding data for antibody Ab3 obtained following the protocol in Example 8 below.

Figure 4 provides human HGF ELISA binding data for antibody Ab4 obtained following the protocol in Example 7, infra.

Figure 5 provides human HGFELISA binding data for antibody Ab5 obtained following the protocol in Example 8, below.

Figure 6 provides the human HGF ELISA binding data for antibody Ab6 obtained following the protocol in Example 8 below.

Figure 7 provides human HGF ELISA binding data for antibody Ab7 obtained following the protocol below.

Figure 8 provides the human HGF ELISA binding data for antibody Ab8 obtained following the protocol in Example 8 below.

Figure 9 provides human HGFELISA binding data for antibody Ab9 obtained following the protocol in Example 8 below.

Figure 10 provides human HGF ELISA binding data for antibody AblO obtained following the protocol in Example 8 below.

Figure 11 provides human HGF ELISA binding data for antibody Abl 1 obtained following the protocol in Example 8 below.

Figure 12 provides human HGF ELISA binding data for antibody Abl2 obtained following the protocol in Example 8 below.

Figure 13 provides human HGFELISA binding data for antibody Abl3 obtained following the protocol in Example 8 below.

Figure 14 provides human HGF ELISA binding data for antibody Abl4 obtained following the protocol in Example 8 below.

Figure 15 provides human HGF ELISA binding data for antibody Abl5 obtained following the protocol in Example 8 below.

Figure 16 provides human HGF ELISA binding data for antibody Abl6 obtained following the protocol in Example 8 below.

Figure 17 provides human HGFELISA binding data for antibody Abl7 obtained following the protocol in Example 8 below.

Figure 18 provides human HGF ELISA binding data for antibody Ab 18 obtained following the protocol in Example 8 below.

Figure 19 provides human HGFELISA binding data for antibody Abl9 obtained following the protocol in Example 8 below.

Figure 20 provides human HGFELISA binding data for antibody Ab20 obtained following the protocol in Example 8, below.

Figure 21 provides human HGFELISA binding data for antibody Ab21 obtained following the protocol in Example 8, below.

Figure 22 provides human HGFELISA binding data for antibody Ab23 obtained following the protocol in Example 8, below.

Figure 23 provides human HGFELISA binding data for antibody Ab24 obtained following the protocol in Example 8, below.

Figure 24 provides human HGFELISA binding data for antibody Ab25 obtained following the protocol in Example 8, below.

Figure 25 provides human HGFELISA binding data for antibody Ab26 obtained following the protocol in Example 8, below.

Figure 26 provides human HGFELISA binding data for antibody Ab27 obtained following the protocol in Example 8, below.

Figure 27 provides human HGFELISA binding data for antibody Ab28 obtained following the protocol in Example 8, below.

Figure 28 presents the subcutaneous U-87MG glioma response to treatment with negative control antibody or AblO or Abl2 (10 mg/kg/injection) obtained following the protocol corresponding to Figures 28 and 29 in Example 10 below.

Figure 29 provides a survival ratio curve for subcutaneous U-87MG glioma treated with negative control antibody or AblO or Abl2 (10 mg/kg/injection) obtained following the protocol for Figures 28 and 29 in Example 10 below.

Figure 30 provides subcutaneous U-87MG gliomas obtained following the protocol corresponding to Figures 30 and 31 as described in Example 10 below, against escalating doses of Ab8 (10, 2.5 and 0.25 mg/kg/injection) or negative Response to control antibody (10 mg/kg/injection) treatment.

Figure 31 provides Ab8 (10, 2.5 and 0.25 mg/kg/injection) or negative control antibody (10 mg/kg/injection) obtained with increasing doses following the experimental protocol described in Example 10 below corresponding to Figures 30 and 31 )-treated subcutaneous U-87MG glioma survival rate curve.

Figure 32 provides subcutaneous U-87MG gliomas obtained following the protocol corresponding to Figures 32 and 33 described in Example 10 below, versus negative Response to control antibody (10 mg/kg/injection) treatment.

Figure 33 provides subcutaneous tissue treated with increasing doses of Ab10 (10, 2.5 and 0.25 mg/kg/injection) or negative control antibody (10 mg/kg/injection) obtained following the protocol corresponding to Figures 32 and 33 in Example 10 below. Survival ratio curve of U-87MG glioma.

Figure 34 provides subcutaneous U-87MG gliomas obtained following the experimental protocol corresponding to Figures 34 and 35 described in Example 10 below, versus escalating doses of Ab28 (30, 10 and 2.5 mg/kg/injection) or Response to negative control antibody (30 mg/kg/injection) treatment.

Figure 35 provides Ab28 (30, 10 and 2.5 mg/kg/injection) or negative control antibody (30 mg/kg/injection) obtained with increasing doses following the experimental protocol corresponding to Figures 34 and 35 described in Example 10 below. )-treated subcutaneous U-87MG glioma survival rate curve.

Figure 36 contains experimental data showing the inhibition of human HGF-driven phosphorylation of Y1234/35, Y1003 and Y1349 of c-met by Ab8 using PC-3 cells (prostate adenocarcinoma) following the protocol described in Example 11 below. change.

Figures 37-50 contain the following experimental results, respectively: Following the experimental protocol in Example 12 below, the analysis of different anti-HGF antibodies (Abl, Ab2, Ab7, Ab8, Ab9, Ab10, Ab12, Ab14, Ab19, Ab21, Ab23, Ab24, Ab25 and Ab28) on the human HGF-driven cell proliferation of 4mBr-5 cells (rhesus monkey bronchial epithelial cells).

Figure 51 contains the results of the following experiment: Analysis of human HGF drive of DBTRG cells (human glioblastoma) by an anti-HGF antibody (Ab8) according to the invention using a Matrigel chamber following the protocol described in Example 13 below The role of cell invasion.

Figures 52A-52G provide the polypeptide sequences of the full-length heavy chains of antibodies Abl-Ab21 and Ab23-28, aligned by their framework regions (FR), complementarity determining regions (CDR) and constant regions.

Figures 53A-53D provide the polypeptide sequences of the full-length light chains of antibodies Abl-Ab21 and Ab23-28, aligned by their framework regions (FR), complementarity determining regions (CDR) and constant regions.

Figures 54A-54S provide the polynucleotide sequences encoding the full-length heavy chains of antibodies Abl-Ab21 and Ab23-28, aligned by their framework regions (FR), complementarity determining regions (CDR) and constant regions.

Figures 55A-55J provide the polynucleotide sequences encoding the full-length light chains of antibodies Abl-Ab21 and Ab23-28, aligned by their framework regions (FR), complementarity determining regions (CDR) and constant regions.

Figure 56 provides the polypeptide sequence coordinates of the variable regions and complementarity determining regions (CDRs) of the heavy chains of antibodies Abl-Ab21 and Ab23-28.

Figure 57 provides polypeptide sequence coordinates for the constant and framework regions (FR) of the heavy chains of antibodies Abl-Ab21 and Ab23-28.

Figure 58 provides the polypeptide sequence coordinates of the variable regions and complementarity determining regions (CDRs) of the light chains of antibodies Abl-Ab21 and Ab23-28.

Figure 59 provides polypeptide sequence coordinates for the constant and framework regions (FR) of the light chains of antibodies Abl-Ab21 and Ab23-28.

Figure 60 provides the polynucleotide sequence coordinates of the variable regions and complementarity determining regions (CDRs) of the heavy chains of antibodies Abl-Ab21 and Ab23-28.

Figure 61 provides the polynucleotide sequence coordinates for the constant and framework regions (FR) of the heavy chains of antibodies Abl-Ab21 and Ab23-28.

Figure 62 provides the polynucleotide sequence coordinates of the variable regions and complementarity determining regions (CDRs) of the light chains of antibodies Abl-Ab21 and Ab23-28.

Figure 63 provides the polynucleotide sequence coordinates for the constant and framework regions (FR) of the light chains of antibodies Abl-Ab21 and Ab23-28.

Detailed Description of the Preferred Embodiment

definition

It is to be understood that this invention is not limited to the particular methodology, protocols, cell lines, animal species or genera and reagents described, as such may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the invention which will be limited only by the appended claims.

As used herein, the singular forms "a/an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells and reference to "the protein" includes reference to one or more proteins and equivalents thereof known to those skilled in the art, and the like. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

Hepatocyte Growth Factor (HGF): HGF as used herein does not only encompass the following preproprotein amino acid sequence available as NCBI Reference Sequence: NP_000592.3 (Homo sapiens HGF isoform 1 preproprotein):

(SEQ ID NO: 1081), and encompasses any original, mature, soluble and/or membrane-bound forms of this HGF amino acid sequence, as well as mutants (mutiens (mutien)), splice variants, isoforms, directly Homologues, homologues and variants.

Hepatocyte Growth Factor Receptor (HGF-R): The terms "HGF-R" and "c-met" as used herein refer to the cellular receptor for Hepatocyte Growth Factor (HGF), which generally includes an extracellular domain, Transmembrane domains and intracellular domains, as well as variants and fragments thereof that retain the ability to bind HGF, and include polypeptide molecules, comprise the full-length native amino acid sequence. Human hepatocyte growth factor (HGF) is a multifunctional heterodimeric polypeptide produced by mesenchymal cells. HGF has been shown to stimulate angiogenesis, morphogenesis, and motility, as well as growth and dispersion of various cell types (Bussolino et al., J. Cell. Biol. 119:629, 1992; Zarnegar and Michalopoulos, J. Cell. Biol. 129: 1177, 1995; Matsumoto et al., Ciba. Found. Symp. 212: 198, 1997; Birchmeier and Gherardi, Trends Cell. Biol. 8: 404, 1998; Xin et al. Am. J. Pathol. 158: 1111, 2001 ). The pleiotropic activity of HGF is mediated by its receptor, a transmembrane tyrosine kinase encoded by the proto-oncogene c-met. In addition to regulating various normal cellular functions, HGF and its receptor c-met have been shown to be involved in tumor initiation, invasion and metastasis (Jeffers et al., J. Mol. Med. 74:505, 1996; Comoglio and Trusolino, J. Clin. Invest. 109:857, 2002). HGF/c-met is coexpressed, often overexpressed, on a variety of human solid tumors, including those derived from lung, colon, rectum, stomach, kidney, ovary, skin, multiple myeloma, and thyroid tissue ( People such as Prat, Int.J.Cancer49:323,1991; People such as Chan, Oncogene2:593,1988; People such as Weidner, Am.J.Respir.Cell.Mol.Biol.8:229,1993; People such as Derksen, Blood 99:1405, 2002). HGF acts as an autocrine (Rong et al., Proc. Natl. Acad. Sci. USA 91:4731, 1994; Koochekpour et al., Cancer Res. 57:5391, 1997) and paracrine growth factor (Weidner et al., Am. J. Respir. Cell. Mol. Biol. 8:229, 1993) and anti-apoptotic regulators (Gao et al., J. Biol. Chem. 276:47257, 2001). HGF is a 102kDa protein whose sequence and structure are similar to plasminogen and other thrombin (Nakamura et al., Nature 342:440, 1989; Weidner et al., Am.J.Respir.Cell.Mol.Biol.8 :229,1993). Human HGF is synthesized as a 728 amino acid precursor (preproHGF) that is cleaved intracellularly to an inert single-chain form (proHGF) (Nakamura et al., Nature 342:440, 1989; Rosen et al., J. Cell. Biol .127:1783, 1994). After extracellular secretion, proHGF is cleaved to produce a biologically active disulfide-linked heterodimeric molecule composed of a subunit and n subunit (Nakamura et al., Nature 342:440, 1989; Naldini et al., EMBOJ.11 :4825,1992). The alpha subunit contains 440 residues (69 kDa glycosylation) and consists of an N-terminal hairpin domain and four kringle domains. The β subunit contains 234 residues (34 kDa) and has a serine protease-like domain that lacks proteolytic activity. HGF has two unique cell-specific binding sites: a high-affinity (Kd=2×10 ^-10 M) binding site for the c-met receptor and a low-affinity (Kd=10 M) binding site for heparan sulfate proteoglycan (HSPG). ^-9 M) binding sites, which are present on the cell surface and on the extracellular matrix (Naldini et al., Oncogene 6:501, 1991; Bardelli et al., J.Biotechnol.37:109, 1994; Sakata et al., J.Biol. Chem., 272:9457, 1997).

"C-met" or "HGF-R" is a member of the class IV protein tyrosine kinase receptor family. The full-length c-met gene was cloned and identified as a c-met proto-oncogene (Cooper et al., Nature 311:29, 1984; Park et al., Proc. Natl. Acad. Sci. USA 84:6379, 1987). NK2 (a protein comprising the N-terminus of the α subunit and the first two kringle domains) is sufficient to bind to c-met and activate the signaling cascade for activity, whereas the full-length protein is required for the mitogenic response (Weidner et al., Am. J Respir. Cell. Mol. Biol. 8:229, 1993). HSPG binds to HGF by interacting with the N-terminus of HGF. HGF/c-met has been reported to play an important role in several aspects of cancer development, such as tumor initiation, invasion, metastasis, regulation of apoptosis and angiogenesis. Several different approaches have been investigated to obtain potent antagonistic molecules: truncated HGF proteins such as NK1 (N-terminal domain plus kringle domain 1; Lokker et al., J. Biol. Chem. 268:17145, 1993), NK2 (N-terminal domain plus kringle domains 1 and 2; Chan et al., Science 254:1382,1991) and NK4 (N-terminal domain plus four kringle domains; Kuba et al., CancerRes.60:6737,2000), anti-c- metmAb (Dodge, Master's Thesis, San Francisco State University, 1998) and anti-HGF mAb (Cao et al., Proc. Natl. Acad. Sci. USA98:7443, 2001).

As used herein, the term "neutralizing or antagonizing anti-HGF antibody or antibody fragment" or "HGF antibody antagonist" refers to a monoclonal antibody (mAb) (i.e., anti-HGF mAb) that binds HGF, wherein the binding partially or completely inhibits the activity of HGF One or more biological activities (ie when using the mAb as a single agent). The biological properties of HGF that can be inhibited by neutralizing antibodies are in particular the ability of HGF to bind to its c-met receptor, thereby causing the dispersion of certain cell lines (such as Madin-Darby canine kidney (MDCK) cells); Stimulates the proliferation (i.e., mitogenic) of certain cells, including hepatocytes, Mv1Lu mink lung epithelial cells, and various human tumor cells; causes specific cell dispersion; stimulates angiogenesis, for example when applied to chicken chorioallantois (CAM ) when measured by stimulating human umbilical vascular endothelial cell (HUVEC) proliferation or tube formation or by inducing blood vessels; promoting cell invasion or metastasis; and promoting fibrosis.

A "blocking" antibody or "antagonist" antibody is preferably an antibody that inhibits or reduces the biological activity of the antigen to which it binds, eg the activated HGFβ chain or site/epitope on c-met to which activated HGFβ binds. Preferably, blocking antibodies or antagonist antibodies substantially or completely inhibit the biological activity of the antigen.

An "agonist antibody" as used herein is an antibody that mimics at least one of the functional activities of a polypeptide of interest (eg, the antibody may provide at least one of the c-met activation functions that activate the HGF beta chain).

A "disorder" is any condition that would benefit from treatment with a substance/molecule or method of the invention. Such conditions include chronic and acute conditions or diseases, including those pathological conditions that predispose a mammal to the condition in question. Non-limiting examples of conditions to be treated herein include malignant and benign tumors; non-leukemic and lymphoid malignancies; neuronal, glial, astrocyte, hypothalamic and other glandular, macrophage, epithelial Cellular, stromal, and blastocoel disorders; and inflammatory, immune, and other angiogenesis-related disorders.

The terms "cell proliferative disorder" and "proliferative disorder" refer to a disorder associated with some degree of abnormal cell proliferation. In one embodiment, the cell proliferative disorder is cancer.

"Tumor" as used herein refers to all neoplastic cell growth and proliferation, whether malignant or benign, and to all precancerous and cancerous cells and tissues. The terms "cancer", "cancerous", "cell proliferative disorder", "proliferative disorder" and "tumor" are not mutually exclusive when referred to herein.

The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is often characterized by unregulated cell growth/proliferation. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. More specific examples of such cancers include squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, peritoneal carcinoma, hepatocellular carcinoma, gastrointestinal cancer, pancreatic cancer, colloid Blastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney cancer, liver cancer, prostate cancer, Vulvar cancer, thyroid cancer, liver cancer and various types of head and neck cancer.

"Treatment" as used herein refers to clinical intervention that seeks to alter the natural course of the individual or cell being treated, and may be performed prophylactically or during the course of clinical pathology. Desirable effects of treatment include prevention of occurrence or recurrence of disease, alleviation of symptoms, attenuation of any direct or indirect pathological consequences of disease, prevention of metastasis, reduction of the rate of disease progression, amelioration or palliation, and remission or improved prognosis. In some embodiments, the antibodies of the invention are used to delay the development of a disease or disorder.

An "effective amount" refers to an amount effective, at dosages and for periods of time required, to achieve the desired therapeutic or prophylactic result. A "therapeutically effective amount" of a substance/molecule (agonist or antagonist) of the present invention may vary depending on factors such as the condition of the individual, age, sex and weight, and the presence of the substance/molecule (agonist or antagonist) in the individual The ability to elicit a desired response. A therapeutically effective amount is also one in which any toxic or detrimental effects of the substance/molecule (agonist or antagonist) are outweighed by the therapeutically beneficial effects.

A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time required, to achieve the desired prophylactic result. Usually, but not necessarily, a prophylactically effective amount will be less than a therapeutically effective amount because prophylactic doses are used in a subject prior to or early in the course of disease.

The term "cytotoxic agent" as used herein refers to a substance that inhibits or prevents the function of cells and/or causes damage to cells. The term is intended to include radioactive isotopes (such as those of At ²¹¹ , I ¹³¹ , I ¹²⁵ , Y ⁹⁰ , Re ¹⁸⁶ , Re ¹⁸⁸ , Sm ¹⁵³ , Bi ²¹² , P ³² , and Lu); chemotherapeutic agents such as methotrexate methotrexate, adriamicin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, Mitomycin C, chlorambucil, daunorubicin or other intercalating agents, enzymes and fragments thereof, such as nucleolytic enzymes, antibiotics and toxins, such as bacteria, fungi, plants Or small molecule toxins or enzyme-active toxins of animal origin, including fragments and/or variants thereof; and various antitumor or anticancer agents disclosed below. Other cytotoxic agents are described below. Tumorcidal agents cause the destruction of tumor cells.

A "chemotherapeutic agent" is a compound useful in the treatment of a proliferative disease such as cancer. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carbosulfan Quinones, meturedopa, and uredopa; ethyleneimines and methylamelamines, including altretamine, triethylenemelamine, triethylenephosphamide (trietylenephosphoramide and trimethylolomelamine); acetogenin (especially bullatacin and bullatacinone); camptothecin (including the synthetic analogue topotecan ( topotecan); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogs ); cryptophycin (specifically cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including synthetic analogs KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyn; spongistatin; nitrogen mustards such as chlorambucil, naphthalene, Chlorphosphamide, estramustine, ifosfamide, methylenedichlorodiethylamine, methylenedichlorodiethylamine oxide hydrochloride, melphalan, new nitrogen mustard (novembichin), cholesteryl phenylacetate mustard (phenesterine), prednimustine, trofosfamide, uracilmustard; nitrosoureas such as carmustine, chlorozotocin ), fotemustine, lomustine, nimustine and ranimustine; antibiotics such as enediyne antibiotics (e.g. calicheamicin , especially calicheamicin γ1I and calicheamicin ωI1 (see for example Agnew, ChemIntl.Ed.Engl., 33:183-186 (1994)); dynemicins, including dynatomycin A; bisphosphonates, such as clodronate; esperamicin; and neocarcinogen chromophores and related chromophores Diacetylene antibiotic chromophore), aclacinomysin, actinomycin, anthramycin, azaserine, bleomycin, actinomycin C (cactinomycin), carabicin, caminomycin, carzinophilin, chromomycin, dactinomycin, daunomycin, detorubicin (detorubicin), 6-diazo-5-oxo-L-norleucine, Doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolinyl-doxorubicin, and deoxydoxorubicin), epirubicin ), esorubicin, idarubicin, marcellomycin, mitomycin (such as mitomycin C), mycophenolic acid, nogamycin nogalamycin, olivomycin, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin , streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolite Drugs such as methotrexate and 5-fluorouracil (5-FU); folate analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purines Analogs, such as fludarabine (fludarabine), 6-mercaptopurine, thiametine, thioguanine; pyrimidine analogs, such as cyclocitidine, azacitidine, 6-azuridine, carmofur ), cytarabine, dideoxyuridine, deoxyfluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolonepropionate, epitiostanol, mepitiostane, testolactone; antiadrenal agents such as aminoglutethimide, mitotane, trilostane; folic acid supplements, Such as folinic acid (frolinic acid); aceglatone (aceglatone); aldophosphamide glycoside (aldophosphamideglycoside); aminolevulinic acid (aminolevulinicacid); eniluracil (eniluracil); amsacrine (amsacrine); bestrabucil); bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids , such as maytansine and ansamitocin; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin (pentostatin); phenamet; pirarubicin; losoxantrone; podophyllinicacid; 2-acetylhydrazide; procarbazine; Polysaccharide complex (JHS Natural Products, Eugene, Oreg.); Razoxane; Rhizoxin; Sizofuran; Spirogermanium; Tenuazonic acid; Triimine quinone (triaziquone); 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurinA, roridinA and anguidine; urethan; vindesine; dacarbazine; mannomustine; dibromomannitol; dibromodulactol; pipobroman; gacytosine; cytarabine ("Ara-C");cyclophosphamide;thiotepa; Paclitaxel (Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE.TM. Paclitaxel-free Cremophor-free albumin-engineered nanoparticle formulation (American Pharmaceutical Partners, Schaumberg, Ill.) and doxetaxel (Rhone-Poulenc Rorer, Antony, France); chlorambucil; Gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone ; vincristine; vinorelbine; novantrone; teniposide; edatrexate; daunorubicin; aminopterin; xeloda; ibandronate ); CPT-11; topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoids, such as retinoic acid; capecitabine (capecitabine); and any of the above A pharmaceutically acceptable salt, acid or derivative of one.

Also included in the definition of "chemotherapeutic agent" above are antihormonal agents that act to modulate or inhibit the effects of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen ( tamoxifen) (including Tamoxifen), raloxifene, droloxifene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, Ona onapristone and FARESTON toremifene; aromatase inhibitors that inhibit aromatase, which regulates estrogen production in the adrenal gland, such as 4(5)-imidazole, aminoglutethimide, megestrol acetate (megestrolacetate), Exemestane, formestanie, fadrozole, Vorozole, letrozole and anastrozole; and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin ); and troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); antisense oligonucleotides, specifically to inhibit signaling pathways involved in abnormal cell proliferation Those antisense oligonucleotides expressed by genes in, such as PKC-α, Ralf and H-Ras; ribonucleases, such as VEGF expression inhibitors (e.g. ribonuclease) and HER2 expression inhibitors; vaccines, such as gene therapy vaccines, e.g. vaccine, vaccines and vaccine; rIL-2; Topoisomerase 1 inhibitors; rmRH; and a pharmaceutically acceptable salt, acid or derivative of any of the above.

"Growth inhibitory agent" as used herein refers to a compound or composition that inhibits the growth of cells that are contingent on HGF/c-met activation in vitro or in vivo. Thus, the growth inhibitory agent may be an inhibitor that significantly reduces the percentage of HGF/c-met-dependent cells in S phase. Examples of growth inhibitory agents include agents that block cell cycle progression (at positions other than S phase), such as agents that induce G1 arrest and M phase arrest. Classical M-phase blockers include vinca (vincristine and vinblastine), taxanes, and topoisomerase II inhibitors such as doxorubicin, epirubicin, daunomycin, etoposide and bleomycin. Those agents that block G1 also penetrate deep into S-phase arrest, such as DNA alkylating agents such as tamoxifen, prednisone, dacarbazine, methylene chloride, cisplatin, methotrexate, 5 - Fluorouracil and cytarabine. Further information can be found in The Molecular Basis of Cancer, edited by Mendelsohn and Israel, Chapter 1, entitled "Cell cycle regulation, oncogenes, and antineoplastic drugs", Murakami et al. (WBSaunders: Philadelphia, 1995), especially page 13. Taxanes (paclitaxel and docetaxel) are anticancer drugs derived from the yew tree. Docetaxel ( Rhone-PoulencRorer) derived from the European yew, for paclitaxel ( Bristol-Myers Squibb) semi-synthetic analog. Paclitaxel and docetaxel promote the assembly of microtubules from tubulin dimers and stabilize microtubules by preventing depolymerization, which contributes to the inhibition of mitosis in cells.

"Doxorubicin" is an anthracycline antibiotic. The complete chemical name of doxorubicin is (8S-cis)-10-[(3-amino-2,3,6-trideoxy-α-L-lyxose-hexopyranosyl)oxy base]-7,-8,9,10-tetrahydro-6,8,11-trihydroxy-8-(hydroxyacetyl)-1-methoxy-5,12-naphthalenedione.

The terms "cell proliferative disorder" and "proliferative disorder" herein refer to a disorder associated with some degree of abnormal cell proliferation. In one embodiment, the cell proliferative disorder is cancer or tumor. As noted previously, "tumor" as used herein refers to all neoplastic cell growth and proliferation, whether malignant or benign, and to all precancerous and cancerous cells and tissues. The terms "cancer", "cancerous", "cell proliferative disorder", "proliferative disorder" and "tumor" are not mutually exclusive when referred to herein.

The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is often characterized by unregulated cell growth/proliferation. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. More specific examples of such cancers include squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, peritoneal carcinoma, hepatocellular carcinoma, gastrointestinal cancer, pancreatic cancer, colloid Blastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney cancer, liver cancer, prostate cancer, Vulvar cancer, thyroid cancer, liver cancer and various types of head and neck cancer.

The term "dysregulation of angiogenesis" herein includes any condition in which abnormal angiogenesis is present. Such conditions include non-neoplastic and neoplastic conditions. Neoplastic conditions include, but are not limited to, the cancers described above. Non-neoplastic conditions include, but are not limited to, undesirable or abnormal hypertrophy, arthritis, rheumatoid arthritis (RA), psoriasis, psoriatic plaques, sarcoidosis, atherosclerosis, atherosclerosis Diabetic and other proliferative retinopathy, including retinopathy of prematurity, retrolentic fibroplasia, neovascular glaucoma, age-related macular degeneration, diabetic macular edema, corneal neovascularization, corneal transplantation neovascularization , corneal graft rejection, retinal/choroidal neovascularization, canthal neovascularization (iris redness), ocular neovascular disease, vascular restenosis, arteriovenous malformation (AVM), meningioma, hemangioma, angiofibroma , thyroid hyperplasia (including Graves' disease), corneal and other tissue transplantation, chronic inflammation, pulmonary inflammation, acute lung injury/ARDS, sepsis, essential pulmonary hypertension, malignant pulmonary effusion, cerebral edema (eg, with acute stroke/closed head injury/trauma-related), synovial inflammation, pannus formation in RA, myositis ossificans, hypertrophic bone formation, osteoarthritis (OA), refractory ascites, polycystic Ovarian disease, endometriosis, 3rd compartment humoral disease (pancreatitis, compartment syndrome, burns, enteropathy), uterine fibroids, premature labor, chronic inflammation such as IBD (Crohn's disease and ulcerative colon inflammation), renal allograft rejection, inflammatory bowel disease, nephrotic syndrome, undesired or abnormal mass growth of tissue (not cancer), haemophilic joints, hypertrophic scarring, inhibited hair growth, Osler- Weber syndrome, pyogenic granulomatoma retrolentic fibroplasia, scleroderma, trachoma, vascular adhesions, synovitis, dermatitis, preeclampsia, ascites, pericardial effusions (such as those associated with pericarditis ) and pleural effusion.

The term "recombinant cell" or "recombinant host cell" herein generally refers to any cell engineered to express one or more antibody polypeptides according to the invention. Such cells include, for example, bacterial, fungal, yeast, mammalian, invertebrate (such as insect), plant and avian cells. Preferred host cells are yeast, fungi, especially filamentous fungi, and mammalian cells. Yeast and filamentous fungi include, but are not limited to, Pichiapastoris, Pichiafinlandica, Pichiatrehalophila, Pichiakoclamae, membrane Pichiamembranaefaciens, Pichiaminuta (Ogataeaminuta, Pichialindneri), Pichiaopuntiae, Pichiathermotolerans ), Pichiasalctaria, Pichiaguercuum, Pichiapijperi, Pichiastiptis, Pichiamethanolica, Pichia Red yeast sp., Saccharomycescerevisiae, Saccharomyces sp., Hansenula polymorpha, Kluyveromyces sp., Kluyveromyceslactis, Candida albicans, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei, Chrysosporium lucknowense, Fusarium sp., Cereal Fusarium gramineum, Fusarium venenatum, Physcomitrellapatens and Neurospora crassa, Pichia sp., Any Saccharomyces sp., Hansenula polymorpha, any Kluyveromyces sp., Candida albicans, any Aspergillus sp., Trichoderma reesei, Chrysosporium luknovensii, any Fusarium sp., and Neurospora crassa.

Examples of invertebrate cells include insect cells, such as Drosophila S2 and Spodoptera Sf9, and plant cells.

Examples of suitable mammalian host cell lines include Chinese hamster ovary (CHO) and COS cells. More specific examples include the SV40-transformed monkey kidney CV1 line (COS-7, ATCCCRL1651); the human embryonic kidney line (subcloned for 293 or 293 cells grown in suspension culture, Graham et al., J. GenVirol. , 36:59(1977)); Chinese hamster ovary cell/-DHFR (CHO, Urlaub and Chasin, Proc.Natl.Acad.Sci.USA, 77:4216(1980)); Mouse Sertolicell (TM4, Mather, Biol. Reprod., 23:243-251 (1980)); human lung cells (W138, ATCCCCL75); human hepatocytes (HepG2, HB8065); and mouse mammary gland tumors (MMT060562, ATCCCCL51). Selection of appropriate host cells is considered to be within the skill of the art. Preferred mammalian cells for antibody expression include CHO cells and COS cells. In an exemplary embodiment, the recombinant host cell is a haploid or polyploid yeast cell of the genus Pichia.

Mating Competent Yeast Species: In the present invention, this is intended to broadly cover any diploid or tetraploid yeast that can grow in culture. The yeast species may exist in haploid, diploid or other polyploid forms. Cells with a given ploid can be propagated in that form for an indefinite number of passages under appropriate conditions. Diploid cells can also spore to form haploid cells. Serial mating can produce tetraploid strains through further mating or fusion of diploid strains. The invention encompasses the use of haploid yeast as well as diploid or other polyploid yeast cells produced, for example, by mating or spheroplast fusion.

Mating-competent yeasts include yeasts that are members of the Saccharomyces family, which includes Arxiozyma; Ascobotryozyma; Citeromyces; Debaryomyces; Debaryomyces; Dekkera; Eremothecium; Issatchenkia; Kazachstania; Kluyveromyces; Kodamaea; Lodderomyces ; Pachysolen; Pichia; Saccharomyces; Saturnispora; Tetrapisispora; Torulaspora; Williopsis; and Zygosaccharomyces. Other types of yeast that may be suitable for use in the present invention include Yarrowia; Rhodosporidium; Candida; Hansenula; ); Sporidiobolus; Bullera; Leucosporidium and Filobasidella.

In a preferred embodiment of the invention, the mating competent yeast is a member of the genus Pichia. In another preferred embodiment of the present invention, the mating-competent Pichia yeast is one of the following species: Pichia pastoris, Pichia methanolica, and Hansenula polymorpha (Pichia angusii). Red yeast (Pichia angusta)). In a particularly preferred embodiment of the invention, the mating competent yeast of the genus Pichia is the species Pichia pastoris.

Haploid Yeast Cell: A cell that has a single copy of each gene of its normal genome (chromosomal) complement.

Polyploid Yeast Cell: A cell that has more than one copy of its normal genome (chromosomal) complement.

Diploid yeast cell: A cell with two copies (alleles) of essentially every gene of its normal genomic complement, usually formed by the process of fusion (mating) of two haploid cells.

Tetraploid Yeast Cell: A cell with essentially four copies (alleles) of each gene of its normal genomic complement, usually formed by the process of fusion (mating) of two haploid cells. Tetraploids can carry two, three, four or more different expression cassettes. The tetraploids can be obtained by selective mating of homoplasmic heterothallic a/a and α/α diploids in Saccharomyces cerevisiae and by serial mating of haploids to obtain auxotrophic diploids in Pichia pastoris obtained from. For example, [methis] haploid can be mated with [adehis] haploid to obtain diploid [his]; and [metarg] haploid can be crossed with [adearg] haploid to obtain diploid [ arg]; followed by diploid [his] × diploid [arg] to obtain tetraploid prototrophic microorganisms. Those skilled in the art will appreciate that references to the benefits and uses of diploid cells also apply to tetraploid cells.

Yeast Mating: The process by which two haploid yeast cells fuse naturally to form a diploid yeast cell.

Meiosis: The process by which a diploid yeast cell undergoes reductional divisions to form four haploid spore products. Each spore can then germinate and form a growing cell line that grows in a haploid fashion.

Selectable marker: A selectable marker is a gene or gene fragment that confers a growth phenotype (a physical growth characteristic) on a cell receiving that gene, eg via a transformation event. A selectable marker allows the cell to survive and grow in a selective growth medium under conditions under which the cell would not be able to grow without receiving the selectable marker gene. Selectable marker genes generally fall into several types, including positive selectable marker genes, such as those that confer resistance to antibiotics or other drugs, temperature (when two temperature-sensitive ("ts") mutants are crossed, or ts mutants are transformed). A gene for resistance; a negative selectable marker gene, such as a biosynthetic gene that confers on cells the ability to grow in a medium that does not contain the specific nutrients required by all cells that do not have that biosynthetic gene, or by not having a wild-type gene A mutagenic biosynthetic gene that confers on cells the ability to fail to grow; and similar genes. Suitable markers include, but are not limited to: ZEO; G418; LYS3; MET1; MET3a; ADE1; ADE3; URA3; and similar markers.

Expression Vectors: These DNA vectors contain elements that facilitate the manipulation of the expression of foreign proteins in the target host cell. Conveniently, manipulation of the sequence and yield of DNA for transformation is first performed in a bacterial host such as E. coli, and the vector will usually include sequences to facilitate this manipulation, including a bacterial origin of replication and an appropriate bacterial selectable marker. Selectable markers encode proteins required for the survival or growth of transformed host cells grown in selective media. Host cells transformed without a vector containing the selection gene will not survive in the medium. Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins, (b) complement auxotrophic deficiencies, or (c) supply critical nutrients not available from complex media. Exemplary vectors and methods for yeast transformation are described, eg, in Burke, D., Dawson, D. and Stearns, T. (2000). Methods in yeast genetics: a Cold Spring Harbor Laboratory course manual. Plainview, N.Y.: Cold Spring Harbor Laboratory Press.

Expression vectors for use in the methods of the invention will also include optional auxotrophic or drug markers for identifying transformed cells such as yeast strains. Drug markers can further be used to amplify the copy number of the vector in host cells.

The sequence encoding the polypeptide of interest is operably linked to transcriptional and translational regulatory sequences that provide for expression of the polypeptide in yeast cells. These vector components may include, but are not limited to, one or more of the following: enhancer elements, promoters, and transcription termination sequences. Sequences for secreting a polypeptide may also include, for example, a signal sequence and the like. The source of replication is optional, since expression vectors typically integrate into the host, eg, the yeast genome. In one embodiment of the invention, the polypeptide of interest is operably linked or fused to a sequence optimized for secretion of the polypeptide from yeast diploid cells.

A nucleic acid is "operably linked" to another nucleic acid sequence when placed into a functional relationship. For example, the DNA of a signal sequence is operably linked to the polypeptide DNA if it is expressed as a preprotein involved in the secretion of the polypeptide; it is operably linked to the coding sequence if a promoter or enhancer affects the transcription of the sequence. Generally, "operably linked" means that the DNA sequences to be linked are contiguous, and, in the case of a secretory leader, contiguous and in reading frame. However, enhancers need not be contiguous. Linking is accomplished by ligation at appropriate restriction sites or via PCR/recombination methods familiar to those skilled in the art (GatewayR Technology; Invitrogen, Carlsbad California). If such sites do not exist, synthetic oligonucleotide linkers or adapters are used according to conventional practice.

A promoter is an untranslated sequence (typically within about 100 to 1000 bp) located upstream (5') of the start codon of a structural gene that controls the transcription and translation of a specific nucleic acid sequence to which it is operably linked. The promoters fall into several categories: inducible, constitutive and repressible promoters (which increase transcription levels in response to the absence of inhibitors). An inducible promoter can cause increased levels of transcription from DNA under its control in response to some change in culture conditions, such as the presence or absence of nutrients or a change in temperature.

The promoter segment can also serve as a site for homologous recombination and integration of the expression vector into the same site in the host genome; or use a selectable marker as a homologous recombination site.

Examples of suitable promoters for Pichia pastoris include the AOX1 promoter (Cregg et al. (1989) Mol. Cell. Biol. 9:1316-1323); the ICL1 promoter (Menendez et al. (2003) Yeast20(13): 1097-108); Glyceraldehyde-3-phosphate dehydrogenase promoter (GAP) (Waterham et al. (1997) Gene186 (1): 37-44); and FLD1 promoter (Shen et al. (1998) Gene216 (1 ):93-102). The GAP promoter is a strong constitutive promoter and the AOX and FLD1 promoters are inducible.

Other yeast promoters include ADH1, alcohol dehydrogenase II, GAL4, PHO3, PHO5, Pyk, and chimeric promoters derived therefrom. Additionally, non-yeast promoters, such as mammalian, insect, plant, reptile, amphibian, bacterial, fungal, viral and avian promoters may be used in the present invention. Most typically, the promoter will comprise a mammalian promoter (which may be endogenous to the gene being expressed) or will comprise a yeast or viral promoter, which provides for efficient transcription in yeast systems.

A polypeptide of interest can be produced not only directly recombinantly, but also as a fusion polypeptide with a heterologous polypeptide, such as a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide. In general, the signal sequence may be a component of the vector, or it may be part of a sequence encoding a polypeptide inserted into the vector. The heterologous signal sequence selected is preferably one that is recognized and processed via one of the standard pathways available within the host cell. The S. cerevisiae alpha factor pre-pro signal has been shown to efficiently secrete various recombinant proteins from Pichia pastoris. Other yeast signal sequences include the alpha mating factor signal sequence, the invertase signal sequence, and signal sequences derived from other secreted yeast polypeptides. In addition, these signal peptide sequences can be engineered to enhance secretion in diploid yeast expression systems. Other secretion signals of interest also include mammalian signal sequences, which may be heterologous to the secreted protein, or which may be the native sequence of the secreted protein. A signal sequence includes a propeptide sequence, and in some cases may include a propeptide sequence. Many such signal sequences are known in the art, including those found on immunoglobulin chains, for example the K28 protoxin sequence, PHA-E, FACE, human MCP-1, human serum albumin signal sequence, human Ig heavy chain, human Ig light chain, etc. See, for example, Hashimoto et al. Protein Eng 11(2) 75 (1998); and Kobayashi et al. Therapeutic Apheresis 2(4) 257 (1998).

Transcription can be increased by inserting transcriptional activator sequences into the vector. These activators are cis-acting elements of DNA, typically about 10 to 300 bp, that act on a promoter to increase its transcription. Transcriptional enhancers are relatively oriented and position-independent within introns as within the coding sequence itself, having been found 5' and 3' of the transcription unit. The enhancer can be spliced into the expression vector at a position 5' or 3' of the coding sequence, but is preferably located at a position 5' from the promoter.

Expression vectors used in eukaryotic host cells may also contain sequences required to terminate transcription and stabilize mRNA. Such sequences are generally available from 3' to the translation termination codon in untranslated regions of eukaryotic or viral DNAs or cDNAs. These regions contain nucleotide segments that are transcribed as polyadenylated fragments in the untranslated portion of the mRNA.

Suitable vectors containing one or more of the components listed above are constructed using standard ligation techniques or PCR/recombination methods. Isolated plasmids or DNA fragments are cleaved, adjusted and rejoined in the form required to generate the desired plasmid or via recombinant methods. For analysis to confirm the correct sequence in the constructed plasmid, the ligation mixture is used to transform host cells and successful transformants are selected by antibiotic resistance (eg ampicillin or Zeocin) where appropriate. Plasmids were prepared from transformants, analyzed by restriction endonuclease digestion and/or sequenced.

As an alternative to restriction and junction fragments, recombination methods based on att sites and recombinases can be used to insert DNA sequences into vectors. Such methods are described, for example, by Landy (1989) Ann. Rev. Biochem. 58:913-949; and are known to those skilled in the art. The method utilizes intermolecular DNA recombination mediated by a mixture of lambda recombinant proteins and recombinant proteins encoding E. coli. Recombination occurs between specific attachment (att) sites on interacting DNA molecules. The att site is described in Weisberg and Landy (1983) Site-Specific Recombination in Phage Lambda, in Lambda II, edited by Weisberg (Cold Spring Harbor, NY: Cold Spring Harbor Press), pp. 211-250. The DNA segments flanking the recombination sites were switched such that after recombination the att sites were a mixed sequence comprising sequences composed of each parental vector. Recombination can occur between DNAs of any topology.

Att sites can be introduced into the sequence of interest by: ligating the sequence of interest into an appropriate vector; through the use of specific primers to generate a PCR product containing the attB site; generating cloning into an appropriate vector containing the att site cDNA library in ; and the like.

Fold as used herein refers to the three-dimensional structure of polypeptides and proteins in which interactions between amino acid residues act to stabilize the structure. While non-covalent interactions are important in determining structure, proteins of interest will typically have intramolecular and/or intermolecular covalent disulfide bonds formed through two cysteine residues. With respect to naturally occurring proteins and polypeptides, or derivatives and variants thereof, proper folding is generally the arrangement that results in optimal biological activity, and can be conveniently monitored by activity assays, such as ligand binding, enzymatic activity, and the like.

In some cases, such as where the desired product is of synthetic origin, assays based on biological activity will be less meaningful. Proper folding of the molecule can be determined based on physical properties, energetic considerations, modeling studies, and the like.

The expression host can be further modified by introducing sequences encoding one or more enzymes that enhance folding and disulfide bond formation, ie, foldases, chaperones, and the like. The sequences can be expressed constitutively or inducibly in yeast host cells using vectors, markers, etc. as known in the art. The sequence includes sufficient transcriptional regulatory elements for the desired expression pattern, preferably stably integrated into the yeast genome via a targeted approach.

For example, eukaryotic PDI is not only an efficient catalyst for protein cysteine oxidation and disulfide bond isomerization, but also exhibits chaperone activity. Coexpression of PDI can promote the production of active proteins with multiple disulfide bonds. Also of interest are BIPs (heavy chain-binding immunoglobulin proteins); cyclophilins; and the like. In one embodiment of the invention, the haploid parent strains each express a different foldase, for example one strain may express BIP while the other strain may express PDI or a combination thereof.

The terms desired protein" or "desired antibody" are used interchangeably and generally refer to a parent antibody or fragment specific for a target (i.e. HGF), or a chimeric or humanized antibody or binding portion thereof derived therefrom, or an antibody or protein containing the same CDRs or epitope specificity as any of the anti-HGF antibodies or fragments described herein.The term "antibody" is intended to include a molecular structure having a specific shape that is suitable for and recognizes an epitope Any polypeptide chain in which one or more non-covalent binding interactions stabilize the complex between the molecular structure and the epitope. The prototype antibody molecule is an immunoglobulin and is derived from all types of immunoglobulins of all origins (IgG , IgM, IgA, IgE, IgD, etc.) are considered "antibodies", such as human, rodent, rabbit, cow, sheep, pig, dog, other mammals, chicken, other poultry, etc. Produce suitable for use as A preferred source of antibodies according to the starting material of the present invention is rabbit. Many sequences encoding antibodies have been described; and others can be produced by methods well known in the art. Examples include chimeric antibodies, human antibodies and other non-human Mammalian antibodies, humanized antibodies, single chain antibodies (such as scFv), camelid antibodies, nanobodies, IgNAR (single chain antibodies derived from sharks), small modular immunopharmaceuticals (SMIPs) and antibody fragments such as Fab, Fab ', F(ab')2, monovalent antibody fragments, such as MetMab-like molecules, IgNar and their analogs. See StreltsovVA et al., Structure of Shark IgNARantibody variable domain and modeling of anearly-development alisotype, Protein Sci. 2005 Nov;14(11):2901-9.版2005年9月30日；GreenbergAS等人,Anewantigenreceptorgenefamilythatundergoesrearrangementandextensivesomaticdiversificationinsharks,Nature.1995年3月9日；374(6518):168-73；NuttallSD等人,Isolationofthenewantigenreceptorfromwobbegongsharks,anduseasascaffoldforthedisplayofproteinlooplibraries,MolImmunol.2001年8月；38(4 ):313-26; Hamers-Casterman C et al., Naturally occurring antibodies devoid of lightchains, Nat 1993 Jun 3;363(6428):446-8; GillDS et al., Biopharmaceutical drug discovery using novel protein scaffolds, Curr Opin Biotechnol. 2006 Dec;17(6):653-8. Electronic 2006 Oct 19.

The invention includes, inter alia, monovalent antibody molecules that bind HGF, similar to the MetMab molecule. MetMab is a specific monovalent antibody to Met. (Met is a protein or fragment thereof encoded by a nucleotide sequence described in Park et al., Proc.Natl.Acad.Sci.84,7479--(1987), and related polypeptides, which include (but are not limited to) allelic variants, splice variants, derivative variants, substitution variants, deletion variants and/or insertion variants, fusion polypeptides and interspecies homologues). The MetMab antibody is a monovalent antibody known under different names (including OA-5d5 (Genentech) and also known as OneArmed5d5, 5d5, MetMab, PRO143966, among others). Antibody OA-5d5, including its structure and properties, and methods of making and using it, are described in US Publication No. 2007/0092520. In one embodiment, an anti-HGF antibody according to the invention may comprise a single Fab region linked to an Fc region. In such embodiments, antibodies of the invention may comprise light chain and heavy chain variable domains as described herein. In such embodiments, the antibody is monovalent and may comprise an entire Fc region. In another such embodiment, the Fc region may comprise at least one bump (knob) and at least one cavity (pore), wherein the presence of the bump and the cavity enhances the relationship between the bump-containing Fc polypeptide and the cavity-containing Fc polypeptide. Complexes are formed between them, for example as described in WO2005/063816. In one embodiment, the Fc region of an antibody of the invention may comprise first and second Fc polypeptides, wherein each of said first and second polypeptides comprises one or more mutations with respect to wild-type human Fc. In one embodiment, the cavity mutation is T366S, L368A and/or Y407V. In another embodiment, the bump mutation is T366W. In a particular embodiment, a monovalent antibody according to the invention may comprise a one-armed antibody synthesized as described in WO2005/063816. In such embodiments, the one-armed antibody may comprise Fc mutations that constitute "knobs" and "pores" as described in WO2005/063816. For example, the hole mutation can be one or more of T366A, L368A, and/or Y407V in the Fc polypeptide, and the cavity mutation can be T366W. The present invention also relates to anti-human HGF monovalent agents that bind to the same HGF epitope and/or compete with anti-HGF antibodies binding to HGF as the antibodies or antibody fragments disclosed herein.

For example, antibodies or antigen-binding fragments can be produced by genetic engineering. In this technique, as in other methods, antibody-producing cells are sensitized to the desired antigen or immunogen. Messenger RNA isolated from antibody-producing cells was used as a template for the preparation of cDNA using PCR amplification. Libraries of vectors are generated by inserting appropriate portions of amplified immunoglobulin cDNAs into expression vectors, each vector containing one heavy chain gene and one light chain gene retaining the original antigen specificity. Combinatorial libraries are constructed by combining a heavy chain gene library with a light chain gene library. This produces a clonal library that co-expresses heavy and light chains (similar to Fab fragments or antigen-binding fragments of antibody molecules). Vectors carrying these genes are co-transfected into host cells. When antibody gene synthesis is induced in a transfected host, heavy and light chain proteins can self-assemble to generate active antibodies, which can be detected by screening with antigen or immunogen.

Contemplated sequences encoding antibodies include those encoded by native sequences, as well as nucleic acids that differ in sequence from the disclosed nucleic acids by virtue of the degeneracy of the genetic code, and variants thereof. Variant polypeptides may include amino acid (aa) substitutions, additions or deletions. Amino acid substitutions can be conservative amino acid substitutions or substitutions that eliminate non-essential amino acids, thereby altering the glycosylation site, or reducing misfolding due to the substitution or deletion of one or more cysteine residues that are not required for function. to minimum. Variants can be designed to retain or enhance the biological activity of specific regions of a protein (eg, functional domains, catalytic amino acid residues, etc.). Variants also include fragments of the polypeptides disclosed herein, in particular biologically active fragments and/or fragments corresponding to functional domains. Techniques for in vitro mutagenesis to clone genes are known. The invention also includes polypeptides that have been modified using ordinary molecular biological techniques in order to improve their resistance to proteolytic degradation or to optimize solubility properties or to make them more suitable as therapeutic agents.

Chimeric antibodies can be produced by recombinant means by combining variable light and heavy chain regions ( _VL and _VH ) obtained from antibody-producing cells of one species with constant light and heavy chain regions from another species . Chimeric antibodies typically utilize rodent or rabbit variable regions and human constant regions in order to produce antibodies with predominantly human domains. The production of such chimeric antibodies is well known in the art and can be accomplished by standard means (as described, for example, in US Patent No. 5,624,659, which is hereby incorporated by reference in its entirety). It is also contemplated that the human constant regions of chimeric antibodies of the invention may be selected from IgG1, IgG2, IgG3 and IgG4 constant regions.

Humanized antibodies are engineered to contain even more human-like immunoglobulin domains and incorporate only the complementarity determining regions of animal-derived antibodies. This is achieved by carefully examining the sequence of the hypervariable loops of the variable regions of monoclonal antibodies and adapting them to the structure of human antibody chains. Although complicated on the surface, the process is actually simple and straightforward. See, eg, US Patent No. 6,187,287, which is incorporated herein by reference in its entirety.

In addition to whole immunoglobulins (or their recombinant counterparts), fragments of immunoglobulins (eg, Fab', F(ab')2, Fab or other fragments) that contain epitope binding sites can be synthesized. "Fragment" or miniature immunoglobulins can be designed using recombinant immunoglobulin technology. For example, "Fv" immunoglobulins for use in the invention can be produced by synthetically fusing a variable light chain region and a variable heavy chain region. Antibody combinations are also of interest, such as diabodies comprising two different Fv specificities. In another embodiment of the invention, immunoglobulin fragments encompass SMIPs (Small Molecule Immunopharmaceuticals), Camelids, Nanobodies and IgNARs.

Immunoglobulins and fragments thereof may be post-translationally modified, e.g., to add effector moieties, such as chemical linkers, and detectable moieties, such as fluorescent dyes, enzymes, toxins, substrates, bioluminescence, may be employed in the methods and compositions of the invention substances, radioactive substances, chemiluminescent moieties and their analogs; or specific binding moieties such as streptavidin, avidin or biotin and their analogs. Examples of additional effector molecules are provided below.

A polynucleotide sequence "corresponds" to a polypeptide sequence if the polynucleotide sequence is translated according to the genetic code to produce the polypeptide sequence (i.e., the polynucleotide sequence "encodes" the polypeptide sequence); An acid sequence "corresponds to" another polynucleotide sequence.

A "heterologous" region or domain of a DNA construct is an identifiable segment of DNA within a larger DNA molecule that is not found related to larger molecules in nature. Thus, when the heterologous region encodes a mammalian gene, the gene will typically be flanked by DNA that does not flank the mammalian genomic DNA in the genome of the source organism. Another example of a heterologous region is a construct in which the coding sequence itself is not found in nature (eg, a cDNA in which the genomic coding sequence contains introns or codons for the synthetic sequence differ from the native gene). Allelic variation or naturally occurring mutational events do not produce heterologous regions of DNA as defined herein.

A "coding sequence" is an in-frame sequence (in terms of the genetic code) of codons corresponding to or encoding a protein or peptide sequence. Two coding sequences, or their complements, encode the same amino acid sequence, then the sequences correspond to each other. A coding sequence, in conjunction with appropriate regulatory sequences, can be transcribed and translated into a polypeptide. A polyadenylation signal and transcription termination sequence will usually be located 3' to the coding sequence. A "promoter sequence" is a DNA regulatory region capable of binding RNA polymerase in a cell and initiating transcription of a downstream (3' direction) coding sequence. Promoter sequences often contain additional sites for the binding of regulatory molecules, such as transcription factors, that affect the transcription of the coding sequence. A coding sequence is under the "control" or "operable" of a promoter sequence when RNA polymerase binds to the promoter sequence in a cell and transcribes the coding sequence into mRNA, which in turn is translated into the protein encoded by the coding sequence ground link" to the promoter.

Vectors are used to introduce foreign substances such as DNA, RNA or proteins into organisms or host cells. Typical vectors include recombinant viruses (for polynucleotides) and liposomes (for polypeptides). A "DNA vector" is a replicon, such as a plasmid, phage, or cosmid, to which another polynucleotide segment can be ligated to cause replication of the ligated segment. An "expression vector" is a DNA vector containing regulatory sequences that will direct synthesis of a polypeptide by an appropriate host cell. This generally means that the promoter binds RNA polymerase and initiates transcription of the mRNA, and the ribosome binding site initiates the signal to direct translation of the mRNA into a polypeptide. The polypeptide encoded by the polynucleotide sequence can be produced by incorporating the polynucleotide sequence into an expression vector at the proper site and in the correct reading frame, and then transforming an appropriate host cell with the vector.

"Amplification" of a polynucleotide sequence is the in vitro production of multiple copies of a particular nucleic acid sequence. The amplified sequence is usually in the form of DNA. Various techniques for performing such amplifications are described in a review paper by Van Brunt (1990, Bio/Technol., 8(4):291-294). The polymerase chain reaction, or PCR, is the prototype of nucleic acid amplification, and the use of PCR herein should be considered as exemplary of other suitable amplification techniques.

The general structure of vertebrate antibodies is now well understood (Edelman, G.M., Ann. N.Y. Acad. Sci., 190:5 (1971)). Antibodies are composed of two identical light polypeptide chains ("light chains") with a molecular weight of approximately 23,000 Daltons and two identical heavy chains ("heavy chains") with a molecular weight of 53,000-70,000. The four chains are connected by disulfide bonds in a "Y" configuration, where the light chain anchor begins at the heavy chain at the opening of the "Y" configuration. The "branch" portion of the "Y" configuration represents the Fab region; the backbone portion of the "Y" configuration represents the FC region. The amino acid sequence is oriented extending from the N-terminal end at the top of the "Y" configuration to the C-terminal end at the bottom of each chain. The N-terminal end has a variable region, which has antigen specificity due to the antigen, and is about 100 amino acids in length, with slight differences between light chains and heavy chains and between antibodies.

In each chain the variable region is linked to a constant region that extends the length of the remaining chain and does not vary with antibody specificity within a particular class of antibody (ie, the antigen is responsible for specificity). There are mainly five classes of known constant regions that determine the class of the heavy chain constant regions of an immunoglobulin molecule (IgG, IgM, IgA, IgD, and IgE correspond to gamma, mu, alpha, delta, and epsilon). The constant regions or classes then determine the effector functions of the antibody, including activation of complement (Kabat, E.A., Structural Concepts in Immunology and Immunochemistry, 2nd ed., pp. 413-436, Holt, Rinehart, Winston (1976)), and other cellular responses (Andrews, D.W. et al., Clinical Immunobiology, pp. 1-18, W.B. Sanders (1980); Kohl, S. et al., Immunology, 48:187 (1983)); whereas the variable region determines the antigen with which it will react. Light chains are classified as either kappa or lambda. Each type of heavy chain can be prepared from either kappa or lambda light chains. The light and heavy chains are covalently bonded to each other, and when the immunoglobulin is produced by a hybridoma or by a B cell, the "tail" portions of the two heavy chains are bonded to each other by a covalent disulfide bond.

The expression "variable region" or "VR" refers to the domains within each pair of light and heavy chains of an antibody that are directly involved in the binding of the antibody to the antigen. Each heavy chain has a variable domain (VH) at one terminus followed by constant domains. Each light chain has a variable domain (VL) at one end and a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain can The variable domains are aligned with the variable domains of the heavy chain.

The expression "complementarity determining region", "hypervariable region" or "CDR" refers to one or more of the hypervariable or complementarity determining regions (CDRs) found in the variable region of the light or heavy chain of an antibody ( See Kabat, E.A. et al., Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, Md., (1987)). These expressions include hypervariable regions or hypervariable loops in the three-dimensional structure of antibodies as defined by Kabat et al. 917 (1987)). The CDRs in each chain are held in close proximity by the framework regions and, together with the CDRs from other chains, contribute to the formation of the antigen binding site. Within the CDRs there are selected amino acids that have been described as selection determining regions (SDRs), which represent key contact residues used by the CDRs in antibody-antigen interactions (Kashmiri, S., Methods, 36:25-34 (2005) ).

An "epitope" or "binding site" is an area or region on an antigen that is specifically bound by an antigen-binding peptide, such as an antibody. A protein epitope may contain amino acid residues that are directly involved in binding (also referred to as the immunodominant component of the epitope) and other amino acid residues that are not directly involved in binding, such as amino acids that can be effectively blocked by specific antigen-binding peptides residues (in other words, the amino acid residues are within the "footprint" of the specific antigen-binding peptide). The term epitope herein includes two types of amino acid binding sites in any specific region of HGF that specifically bind to an anti-HGF antibody. HGF may comprise a variety of different epitopes, which may include, but are not limited to, (1) linear peptide epitopes, (2) conformational epitopes consisting of one or more non-contiguous epitopes that are positioned close to each other in the mature HGF conformation amino acid composition; and (3) post-translational epitopes, which consist entirely or partially of molecular structures, such as carbohydrate groups, covalently linked to the HGF protein.

The phrase that the first antibody "substantially" or "at least partially" binds to the same epitope as the second antibody means that the epitope binding site of the first antibody comprises at least part of the antigen comprising the epitope binding site of the second antibody. 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more of the amino acid residues. Furthermore, that the first antibody substantially or partially binds to the same or overlapping epitope as the second antibody means that the first and second antibodies compete in binding to the antigen, as described above. Thus, the term "binds to substantially the same epitope or determinant as a monoclonal antibody" means that an antibody "competes" with said antibody.

The phrase "binds to the same or overlapping epitope or determinant as the antibody of interest" means that an antibody "competes" with said antibody of interest for at least one or all residues. Identification of one or more antibodies that bind to substantially or substantially the same epitope as the monoclonal antibodies described herein can be readily determined using any of a variety of immunoscreening assays that can assess antibody competition. A variety of such assays are routinely practiced and well known in the art (see, eg, US Patent No. 5,660,827, issued August 26, 1997, which is specifically incorporated herein by reference). It will be appreciated that it is not in any way necessary to actually determine the epitope to which an antibody described herein binds to identify an antibody that binds to the same or substantially the same or overlapping epitope as a monoclonal antibody described herein.

For example, when the test antibody to be examined is obtained from a different animal source, or even has a different Ig isotype, a simple competition assay can be used in which a control antibody is mixed with the test antibody and then applied to a sample containing HGF. ELISA-based protocols, radioimmunoassays, Western blots and analysis using surface plasmon resonance (by instruments such as Biacore) or biofilm layer interferometry (by instruments such as Octet) are suitable for such simple competition studies.

In certain embodiments, a control anti-HGF antibody can be premixed (eg, at a ratio of about 1:1, 1:2, 1:10, or about 1:100) with varying amounts of a test antibody for a period of time and then applied to HGF antigen sample. In other embodiments, the control antibody and varying amounts of the test antibody can simply be added separately and mixed during exposure to the HGF antigen sample. As long as bound antibody can be distinguished from free antibody (e.g. by using separation or washing techniques to eliminate unbound antibody) and control antibody from test antibody (e.g. by using species-specific or isotype-specific secondary antibodies or by using detectable labels Specifically labeling the control antibody), it will be possible to determine whether the test antibody reduces the binding of the control antibody to the HGF antigen, indicating that the test antibody recognizes substantially the same epitope as the control anti-HGF antibody. Binding of a (labeled) control antibody in the presence of a completely irrelevant antibody (antibody that does not bind HGF) can serve as a high control value. A low control value can be obtained by incubating a labeled control antibody with the same but unlabeled control antibody, where competition will occur and binding of the labeled antibody will decrease. A marked decrease in labeled antibody reactivity in the presence of the test antibody in the test assay indicates that the test antibody recognizes substantially the same epitope, ie, the test antibody competes with the labeled control antibody. For example, at any ratio of control antibody:test antibody between about 1:1 or 1:10 and about 1:100, the binding of the control antibody to HGF is reduced by at least about 50%, such as at least about 60%. ) or more, preferably at least about 70% (eg, about 65%-100%) of any test antibody is considered an antibody that binds to substantially the same or overlapping epitope or determinant as the control antibody.

Preferably, such test antibodies will reduce the binding of the control antibody to the HGF antigen by preferably at least about 50%, at least about 60%, at least about 80%, or at least About 90% (eg, about 95%).

Competition can alternatively also be assessed by eg flow cytometry assays. In such tests, HGF-bearing cells can be first incubated with a control antibody that binds HGF, and then incubated with a fluorochrome or biotinylated test antibody. If the binding obtained after preincubation with a saturating amount of the control antibody is about 80% of the binding (as measured by means of fluorescence) obtained by the test antibody without preincubation with the control antibody, preferably about 50%, about 40% % or less (eg, about 30%), the antibody is said to compete with a control antibody. Alternatively, if the binding obtained with a labeled control antibody (via a fluorochrome or biotin) to cells pre-incubated with a saturating amount of the test antibody is about 80% of the binding obtained without pre-incubation with the test antibody, Preferably about 50%, about 40% or less (eg about 30%), the antibody is said to compete with a control antibody.

A simple competition assay can also be advantageously employed, in which the test antibody is preabsorbed at a saturating concentration and applied to the surface on which HGF is immobilized. The surface in simple competition assays is preferably a BIACORE chip (or other medium suitable for surface plasmon resonance analysis). Binding of HGF-binding control antibodies to HGF-coated surfaces was measured. This binding of the control antibody alone to the HGF-containing surface is compared to the binding of the control antibody in the presence of the test antibody. A significant reduction in binding of the control antibody to the HGF-containing surface in the presence of the test antibody indicates that the test antibody recognizes substantially the same epitope as the control antibody and thus "competes" with the control antibody. Any test antibody that reduces binding of the control antibody by at least about 20% or more, at least about 40%, at least about 50%, at least about 70% or more is considered to bind to substantially the same epitope or determinant as the control antibody. cluster antibodies. Preferably, such test antibodies will reduce the binding of the control antibody to HGF by at least about 50% (eg, at least about 60%, at least about 70%, or more). It is understood that the order of the control antibody and the test antibody can be reversed; that is, in a competition assay, the control antibody can be bound to the surface first, and the test antibody subsequently introduced into contact with the surface. Preferably, the antibody with higher affinity for the HGF antigen binds first to the HGF-containing surface, as would be expected, as would be expected to see that binding of the second antibody (assuming the antibody is competing) would be substantially reduced. Additional examples of such assays are provided, eg, in Saunal and Regenmortel, (1995) J. Immunol. Methods 183:33-41, the disclosure of which is incorporated herein by reference.

Also, in particular, Western blot-based assays can be used to determine whether an antibody binds the same or overlapping epitope on HGF as another antibody or an epitope to which a test antibody binds. In this assay, a library of peptides corresponding to the antigen to which the antibody binds (here HGF) is prepared, corresponding to overlapping portions of proteins typically 10-25, 10-20, or 10-15 amino acids long. These different overlapping amino acid peptides covering the HGF sequence were synthesized and covalently bound to PepSpots nitrocellulose membranes (JPT Peptide technologies, Berlin, Germany). Blots were then prepared and probed according to the manufacturer's recommendations.

Essentially, the immunoblot assay then detects by fluorescent means which peptides in the library bind to the test antibody and thereby can identify which residues on the antigen (ie HGF) interact with the test antibody. (See an embodiment of this technology in US Patent No. 7,935,340, which is incorporated herein by reference).

The expression "framework region" or "FR" refers to one or more of the framework regions within the variable regions of antibody light and heavy chains (see Kabat, E.A. et al., Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, Md., (1987 )). These expressions include those regions of amino acid sequence inserted between the CDRs within the variable regions of the antibody light and heavy chains.

Anti-HGF antibodies and binding fragments thereof

Antibody Ab1

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSVEESGGRLVTPGTPLTLTCTVSGFSLSAYAMSWVRQAPEKGLEWIAVIYVIGATDYASWAKGRFTISRTSTTVDLRIPSPTTEDTATYFCARVYDSVWNHFNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:1)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSVEESGGRLVTPGTPLTLTCTVSGFSLSAYAMSWVRQAPEKGLEWIAVIYVIGATDYASWAKGRFTISRTSTTVDLRIPSPTTEDTATYFCARVYDSVWNHFNLWGPGTLVTVSS (SEQ ID NO: 2).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab1相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 10).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：AYDMTQTPASVEVAVGGTVTIKCQASQSISSWLAWYQQKPGQPPKLLIYQASKLASGVPSRFKGSGSGTEFTLTISGVECADAATYYCQQAYSVSNVDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:21)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: AYDMTQTPASVEVAVGGTVTIKCQASQSISSWLAWYQQKPGQPPKLLIYQASKLASGVPSRFKGSGSGTEFTLTISGVECADAATYYCQQAYSVSNVDNAFGGGTEVVVKR (SEQ ID NO: 22).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab1 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 30).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO: 4, SEQ ID NO: 6 and SEQ ID NO: 8, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQ ID NO: 1 or it contains the variable heavy chain sequence of SEQ ID NO: 2; and/or it further contains SEQ ID NO: 24, SEQ ID NO: 26 and SEQ ID NO: One, two or three of the polypeptide sequences of 28, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 21 or the variable light chain sequence containing SEQ ID NO: 22; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7 and SEQ ID NO:9, which correspond to SEQ ID NO: The heavy chain sequence of 1 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:2; And/or one of the polypeptide sequences of SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27 and SEQ ID NO:29 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 21 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 22; or these polypeptide sequences and at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 1 or SEQ ID NO: 2 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 21 or SEQ ID NO: 22 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 4, SEQ ID NO: 6 and SEQ ID NO: 8 or three, which correspond to the heavy chain sequence of SEQ ID NO: 1 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 2; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 24, SEQ ID NO: 26 and SEQ ID NO: 28 or three, which correspond to the light chain sequence of SEQ ID NO: 21 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO: 22; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7 and SEQ ID NO:9 Either, both, three or four, it corresponds to the heavy chain sequence of SEQ ID NO: 1 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 2; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27 and SEQ ID NO: 29 One, two, three or four, which correspond to the light chain sequence of SEQ ID NO: 21 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 22; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: The variable heavy chain region; the variable light chain region of SEQ ID NO: 22; the complementarity determining region (SEQ ID NO: 4, SEQ ID NO: 6 and SEQ ID NO: 8) of the variable heavy chain region of SEQ ID NO: 2; and the variable of SEQ ID NO: 22 the complementarity determining region of the light chain region (SEQ ID NO: 24, SEQ ID NO: 26 and SEQ ID NO: 28); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: Variable heavy chain region; variable light chain region of SEQ ID NO: 22; framework region (SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7 and SEQ ID NO: 9) of the variable heavy chain region of SEQ ID NO: 2; and SEQ ID NO: 22 The framework region of the variable light chain region (SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27 and SEQ ID NO: 29).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab1 comprising or consisting of SEQ ID NO: 1 and SEQ ID NO: 21, or comprising the CDRs of Ab1 and having at least one of the biological activities set forth herein Or an antibody fragment, or an anti-HGF antibody that competes with Ab1 for binding to HGF, preferably an antibody that contains a sequence at least 90% or 95% identical to the sequence of Ab1 or an antibody that binds to the same or overlapping epitope on HGF with Ab1.

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Abl, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 2 and the variable light chain sequence of SEQ ID NO: 22 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:2 and/or SEQ ID NO:22 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl. In another embodiment of the invention, anti-HGF antibodies such as Abl or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab1 and the FR, CDR, variable heavy chain as described above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab2

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：EVQLVESGGGLVQPGGSLRLSCAASGFTVSAYAMSWVRQAPGKGLEWVAVIYVIGATDYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVYDSVWNHFNLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:41)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: EVQLVESGGGLVQPGGSLRLSCAASGFTVSAYAMSWVRQAPGKGLEWVAVIYVIGATDYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVYDSVWNHFNLWGQGTLVTVSS (SEQ ID NO: 42).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab2相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 50).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：DIQMTQSPSTLSASVGDRVTITCQASQSISSWLAWYQQKPGKAPKLLIYQASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQAYSVSNVDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:61)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: DIQMTQSPSTLSASVGDRVTITCQASQSISSWLAWYQQKPGKAPKLLIYQASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQAYSVSNVDNAFGGGTKVEIKR (SEQ ID NO: 62).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab2 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 70).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:44, SEQ ID NO:46 and SEQ ID NO:48, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQ ID NO: 41 or it contains the variable heavy chain sequence of SEQ ID NO: 42; and/or it further contains SEQ ID NO: 64, SEQ ID NO: 66 and SEQ ID NO: One, two or three of the polypeptide sequences of 68, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 61 or the variable light chain sequence containing SEQ ID NO: 62; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47 and SEQ ID NO:49, which correspond to SEQ ID NO: A heavy chain sequence of 41 or a framework region (FR or constant region) of a variable heavy chain sequence of SEQ ID NO:42; and/or one of the polypeptide sequences of SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67 and SEQ ID NO:69 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 61 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 62; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 41 or SEQ ID NO: 42 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 61 or SEQ ID NO: 62 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:44, SEQ ID NO:46 and SEQ ID NO:48 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 41 or the variable heavy chain sequence of SEQ ID NO: 42; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:64, SEQ ID NO:66 and SEQ ID NO:68 or three, which correspond to the light chain sequence of SEQ ID NO: 61 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO: 62; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47 and SEQ ID NO: 49 Either, both, three or four, it corresponds to the heavy chain sequence of SEQ ID NO: 41 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 42; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67 and SEQ ID NO:69 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:61 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:62; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: The variable heavy chain region; the variable light chain region of SEQ ID NO:62; the complementarity determining region (SEQ ID NO:44, SEQ ID NO:46 and SEQ ID NO:48) of the variable heavy chain region of SEQ ID NO:42; and the variable of SEQ ID NO:62 the complementarity determining region of the light chain region (SEQ ID NO:64, SEQ ID NO:66 and SEQ ID NO:68); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: The variable heavy chain region; the variable light chain region of SEQ ID NO:62; the framework region of the variable heavy chain region of SEQ ID NO:42 (SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47 and SEQ ID NO:49); and SEQ ID NO:62 The framework region of the variable light chain region (SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67 and SEQ ID NO: 69).

In a particularly preferred embodiment of the invention, the anti-HGF antibody is Ab2 comprising or consisting of SEQ ID NO: 41 and SEQ ID NO: 61, or an antibody comprising the CDRs of Ab2 and having at least one of the biological activities set forth herein Or an antibody fragment, or an anti-HGF antibody that competes with Ab2 for binding to HGF, preferably an antibody that contains at least 90% or 95% of the same sequence as Ab2, or an antibody that binds to the same or overlapping epitope on HGF with Ab2.

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Ab2, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 42 and the variable light chain sequence of SEQ ID NO: 62 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:42 and/or SEQ ID NO:62 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab2. In another embodiment of the present invention, anti-HGF antibodies such as Ab2 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab2 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab3

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSVEESGGRLVTPGTPLTLTCTVSGLTISSYYMSWVRQAPGKGLEWIGTINPGANTYFASWAKGRFTISRTSTTVDLKITSPTTEDTATYFCAREGDSNDWGVFDLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:81)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSVEESGGRLVTPGTPLTLTCTVSGLTISSYYMSWVRQAPGKGLEWIGTINPGANTYFASWAKGRFTISRTSTTVDLKITSPTTEDTATYFCAREGDSNDWGVFDLWGQGTLVTVSS (SEQ ID NO: 82).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab3相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 90).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：AYDMTQTPASVEIAVGGTVTIRCQASEDIESYLAWYQQKPGQPPKLLIYRASDLASGVSSRFKGSGSGTDYTLTISGVECDDAATYYCQQGYTIDNVDNTFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:101)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: AYDMTQTPASVEIAVGGTVTIRCQASEDIESYLAWYQQKPGQPPKLLIYRASDLASGVSSRFKGSGSGTDYTLTISGVECDDAATYYCQQGYTIDNVDNTFGGGTEVVVKR (SEQ ID NO: 102).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab3 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 110).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:84, SEQ ID NO:86 and SEQ ID NO:88, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQ ID NO: 81 or it contains the variable heavy chain sequence of SEQ ID NO: 82; and/or it further contains SEQ ID NO: 104, SEQ ID NO: 106 and SEQ ID NO: One, two or three of the polypeptide sequences of 108, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 101 or the variable light chain sequence containing SEQ ID NO: 102; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:83, SEQ ID NO:85, SEQ ID NO:87 and SEQ ID NO:89, which correspond to SEQ ID NO: The heavy chain sequence of 81 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:82; And/or one of the polypeptide sequences of SEQ ID NO:103, SEQ ID NO:105, SEQ ID NO:107 and SEQ ID NO:109 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 101 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 102; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 81 or SEQ ID NO: 82 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 101 or SEQ ID NO: 102 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:84, SEQ ID NO:86 and SEQ ID NO:88 or three, which correspond to the heavy chain sequence of SEQ ID NO: 81 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 82; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 104, SEQ ID NO: 106 and SEQ ID NO: 108 or three, which correspond to the light chain sequence of SEQ ID NO: 101 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO: 102; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO:83, SEQ ID NO:85, SEQ ID NO:87 and SEQ ID NO:89 Either, both, three or four, it corresponds to the heavy chain sequence of SEQ ID NO: 81 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 82; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107 and SEQ ID NO: 109 One, two, three or four, which correspond to the light chain sequence of SEQIDNO: 101 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO: 102; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: The variable heavy chain region; the variable light chain region of SEQ ID NO: 102; the complementarity determining region (SEQ ID NO: 84, SEQ ID NO: 86 and SEQ ID NO: 88) of the variable heavy chain region of SEQ ID NO: 82; and the variable of SEQ ID NO: 102 the complementarity determining region of the light chain region (SEQ ID NO: 104, SEQ ID NO: 106 and SEQ ID NO: 108); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: variable heavy chain region; variable light chain region of SEQ ID NO: 102; framework region (SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87 and SEQ ID NO: 89) of the variable heavy chain region of SEQ ID NO: 82; and SEQ ID NO: 102 The framework region of the variable light chain region (SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107 and SEQ ID NO: 109).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab3 comprising or consisting of SEQ ID NO: 81 and SEQ ID NO: 101; or an antibody comprising the CDRs of Ab3 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab3 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab3; or an antibody that binds to an epitope on HGF that is identical or overlapping with Ab3 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Concerning antibody Ab3, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 82 and the variable light chain sequence of SEQ ID NO: 102 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO: 82 and/or SEQ ID NO: 102, which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab3. In another embodiment of the present invention, anti-HGF antibodies such as Ab3 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab3 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab4

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSLEESGGRLVQPGTPLTLSCTASGLTISSYYMSWVRQAPGKGLEWVGTINPGANTYFASSAKGRFTISRSSTTLDLKMTSPTAEDTATYYCAREGDSNDWGVFDLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:121)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSLEESGGRLVQPGTPLTLSCTASGLTISSYYMSWVRQAPGKGLEWVGTINPGANTYFASSAKGRFTISRSSTTLDLKMTSPTAEDTATYYCAREGDSNDWGVFDLWGQGTLVTVSS (SEQ ID NO: 122).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab4相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 130).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：AYDMTQSPASVEAAVGGTVTIRCQASEDIESYLAWYQQKPGQPPKLLIYRASDLASGVSSRFKGSGSGTDYTLTISGLEPEDAATYYCQQGYTIDNVDNTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:141)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: AYDMTQSPASVEAAVGGTVTIRCQASEDIESYLAWYQQKPGQPPKLLIYRASDLASGVSSRFKGSGSGTDYTLTISGLEPEDAATYYCQQGYTIDNVDNTFGGGTKVEIKR (SEQ ID NO: 142).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab4 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 150).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO: 124, SEQ ID NO: 126 and SEQ ID NO: 128, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQ ID NO: 121 or it contains the variable heavy chain sequence of SEQ ID NO: 122; and/or it further contains SEQ ID NO: 144, SEQ ID NO: 146 and SEQ ID NO: One, two or three of the polypeptide sequences of 148, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 141 or the variable light chain sequence containing SEQ ID NO: 142; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127 and SEQ ID NO: 129, which correspond to SEQ ID NO: The heavy chain sequence of 121 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:122; And/or one of the polypeptide sequences of SEQ ID NO:143, SEQ ID NO:145, SEQ ID NO:147 and SEQ ID NO:149 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 141 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 142; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 121 or SEQ ID NO: 122 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 141 or SEQ ID NO: 142 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 124, SEQ ID NO: 126 and SEQ ID NO: 128 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 121 or the variable heavy chain sequence of SEQ ID NO: 122; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 144, SEQ ID NO: 146 and SEQ ID NO: 148 or three, which correspond to the light chain sequence of SEQ ID NO: 141 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO: 142; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127 and SEQ ID NO: 129 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO: 121 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO: 122; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147 and SEQ ID NO: 149 One, two, three or four, which correspond to the light chain sequence of SEQIDNO: 141 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO: 142; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: variable heavy chain region; the variable light chain region of SEQ ID NO: 142; the complementarity determining region (SEQ ID NO: 124, SEQ ID NO: 126 and SEQ ID NO: 128) of the variable heavy chain region of SEQ ID NO: 122; and the variable of SEQ ID NO: 142 the complementarity determining region of the light chain region (SEQ ID NO: 144, SEQ ID NO: 146 and SEQ ID NO: 148); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: variable heavy chain region; variable light chain region of SEQ ID NO: 142; framework region (SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127 and SEQ ID NO: 129) of the variable heavy chain region of SEQ ID NO: 122; and SEQ ID NO: 142 The framework region of the variable light chain region (SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147 and SEQ ID NO: 149).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab4 comprising or consisting of SEQ ID NO: 121 and SEQ ID NO: 141; or an antibody comprising the CDRs of Ab4 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab4 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab4; or an antibody that binds to an epitope on HGF that is identical or overlapping with Ab4 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Concerning antibody Ab4, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 122 and the variable light chain sequence of SEQ ID NO: 142 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO: 122 and/or SEQ ID NO: 142 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab4. In another embodiment of the present invention, anti-HGF antibodies such as Ab4 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab4 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab5

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSLEESGGRLVTPGTPLTLTCTVSGFSLNNYAVGWVRQAPGKGLEWIGIIYLSGNTDYANWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARKFDTGYDIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:161)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSLEESGGRLVTPGTPLTLTCTVSGFSLNNYAVGWVRQAPGKGLEWIGIIYLSGNTDYANWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARKFDTGYDIWGPGTLVTVSS (SEQ ID NO: 162).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab5相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 170).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：AYDMTQTPASMEVAVGGTVTIKCQASQSISTYLAWYQQKPGQPPKLLIYDASDLASGVSSRFKGSGSGTQFTLTISGVECDDAATYYCQQDWSDSNVDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:181)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: AYDMTQTPASMEVAVGGTVTIKCQASQSISTYLAWYQQKPGQPPKLLIYDASDLASGVSSRFKGSGSGTQFTLTISGVECDDAATYYCQQDWSDSNVDNAFGGGTEVVVKR (SEQ ID NO: 182).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab5 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 190).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO: 164, SEQ ID NO: 166 and SEQ ID NO: 168, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO: 161 or it contains the variable heavy chain sequence of SEQIDNO: 162; and/or it further contains SEQIDNO: 184, SEQIDNO: 186 and SEQIDNO: One, two or three of the polypeptide sequences of 188, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 181 or the variable light chain sequence containing SEQ ID NO: 182; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167 and SEQ ID NO: 169, which correspond to SEQ ID NO: the heavy chain sequence of 161 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 162; and/or one of the polypeptide sequences of SEQ ID NO: 183, SEQ ID NO: 185, SEQ ID NO: 187 and SEQ ID NO: 189 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 181 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 182; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 161 or SEQ ID NO: 162 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 181 or SEQ ID NO: 182 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 164, SEQ ID NO: 166 and SEQ ID NO: 168 or three, which correspond to the heavy chain sequence of SEQ ID NO: 161 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 162; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 184, SEQ ID NO: 186 and SEQ ID NO: 188 or three, which correspond to the light chain sequence of SEQ ID NO: 181 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO: 182; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167 and SEQ ID NO: 169 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO: 161 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO: 162; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 183, SEQ ID NO: 185, SEQ ID NO: 187 and SEQ ID NO: 189 One, two, three or four, which correspond to the light chain sequence of SEQIDNO: 181 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO: 182; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: of SEQ ID NO: 162 variable heavy chain region; the variable light chain region of SEQ ID NO: 182; the complementarity determining region (SEQ ID NO: 164, SEQ ID NO: 166 and SEQ ID NO: 168) of the variable heavy chain region of SEQ ID NO: 162; and the variable of SEQ ID NO: 182 the complementarity determining region of the light chain region (SEQ ID NO: 184, SEQ ID NO: 186 and SEQ ID NO: 188); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: of SEQ ID NO: 162 variable heavy chain region; variable light chain region of SEQ ID NO: 182; framework region (SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167 and SEQ ID NO: 169) of the variable heavy chain region of SEQ ID NO: 162; and SEQ ID NO: 182 The framework region of the variable light chain region (SEQ ID NO: 183, SEQ ID NO: 185, SEQ ID NO: 187 and SEQ ID NO: 189).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab5 comprising or consisting of SEQ ID NO: 161 and SEQ ID NO: 181; or an antibody comprising the CDRs of Ab5 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab5 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab5; or an antibody that binds to an epitope on HGF that is identical or overlapping with Ab5 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Ab5, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 162 and the variable light chain sequence of SEQ ID NO: 182 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO: 162 and/or SEQ ID NO: 182, which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab5. In another embodiment of the present invention, anti-HGF antibodies such as Ab5 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab5 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab6

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSVEESGGRLVMPGTPLTLTCTVSGFSLSSNAISWVRQAPEKGLEWIGVIYVIGVTDYASWAQGRFTISKTSTTVDLKIPSPTTEDTATYFCARVYDSGWNHFNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:201)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSVEESGGRLVMPGTPLTLTCTVSGFSLSSNAISWVRQAPEKGLEWIGVIYVIGVTDYASWAQGRFTISKTSTTVDLKIPSPTTEDTATYFCARVYDSGWNHFNLWGPGTLVTVSS (SEQ ID NO: 202).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab6相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 210).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：ADIVMTQTPSSVEAAVGGTVTIKCQASENIYRLLAWYQQKPGQRPKLLIYSASTLASGVPSRFKGSGSGTQFTLTISDLECADAATYYCQNYYYSSRSSYDTYNVFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:221)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: ADIVMTQTPSSVEAAVGGTVTIKCQASENIYRLLAWYQQKPGQRPKLLIYSASTLASGVPSRFKGSGSGTQFTLTISDLECADAATYYCQNYYYSSRSSYDTYNVFGGGTEVVVKR (SEQ ID NO: 222).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab6 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 230).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO: 204, SEQ ID NO: 206 and SEQ ID NO: 208, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO:201 or it contains the variable heavy chain sequence of SEQIDNO:202; And/it further contains SEQIDNO:224, SEQIDNO:226 and SEQIDNO: One, two or three of the polypeptide sequences of 228, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 221 or the variable light chain sequence containing SEQ ID NO: 222; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:203, SEQ ID NO:205, SEQ ID NO:207 and SEQ ID NO:209, which correspond to SEQ ID NO: The heavy chain sequence of 201 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:202; And/or one of the polypeptide sequences of SEQ ID NO:223, SEQ ID NO:225, SEQ ID NO:227 and SEQ ID NO:229 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 221 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 222; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 201 or SEQ ID NO: 202 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 221 or SEQ ID NO: 222 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 204, SEQ ID NO: 206 and SEQ ID NO: 208 or three, which correspond to the heavy chain sequence of SEQ ID NO: 201 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 202; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 224, SEQ ID NO: 226 and SEQ ID NO: 228 or three, which correspond to the light chain sequence of SEQ ID NO: 221 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO: 222; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 203, SEQ ID NO: 205, SEQ ID NO: 207 and SEQ ID NO: 209 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO: 201 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO: 202; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 223, SEQ ID NO: 225, SEQ ID NO: 227 and SEQ ID NO: 229 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:221 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:222; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: The variable heavy chain region; the variable light chain region of SEQ ID NO: 222; the complementarity determining region (SEQ ID NO: 204, SEQ ID NO: 206 and SEQ ID NO: 208) of the variable heavy chain region of SEQ ID NO: 202; and the variable of SEQ ID NO: 222 the complementarity determining region of the light chain region (SEQ ID NO: 224, SEQ ID NO: 226 and SEQ ID NO: 228); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: Variable heavy chain region; the variable light chain region of SEQ ID NO:222; the framework region (SEQ ID NO:203, SEQ ID NO:205, SEQ ID NO:207 and SEQ ID NO:209) of the variable heavy chain region of SEQ ID NO:202; Variable light chain regions and framework regions (SEQ ID NO: 223, SEQ ID NO: 225, SEQ ID NO: 227 and SEQ ID NO: 229).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab6 comprising or consisting of SEQ ID NO: 201 and SEQ ID NO: 221; or an antibody comprising the CDRs of Ab6 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab6 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab6; or an antibody that binds to an epitope on HGF that is identical or overlapping with Ab6 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Concerning antibody Ab6, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 202 and the variable light chain sequence of SEQ ID NO: 222 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO: 202 and/or SEQ ID NO: 222 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab6. In another embodiment of the present invention, anti-HGF antibodies such as Ab6 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab6 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab7

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSVEESGGRLVMPGTPLTLTCTVSGFSLSSNAISWVRQAPEKGLEWIGVIYVIGVTDYASWAQGRFTISKTSTTVDLKIPSPTTEDTATYFCARVYDSGWNHFNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:241)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSVEESGGRLVMPGTPLTLTCTVSGFSLSSNAISWVRQAPEKGLEWIGVIYVIGVTDYASWAQGRFTISKTSTTVDLKIPSPTTEDTATYFCARVYDSGWNHFNLWGPGTLVTVSS (SEQ ID NO: 242).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab7相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 250).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：AYDMTQTPASVEVAVGGTVTIKCQASQSISSWLAWYQQKPGQPPKLLIYEASKLASGVPSRFSGSGSGTQFTLTISGVECADAATYYCQQAYSVANVDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:261)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: AYDMTQTPASVEVAVGGTVTIKCQASQSISSWLAWYQQKPGQPPKLLIYEASKLASGVPSRFSGSGSGTQFTLTISGVECADAATYYCQQAYSVANVDNAFGGGTEVVVKR (SEQ ID NO: 262).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab7 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 270).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:244, SEQ ID NO:246 and SEQ ID NO:248, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO:241 or it contains the variable heavy chain sequence of SEQIDNO:242; And/it further contains SEQIDNO:264, SEQIDNO:266 and SEQIDNO: One, two or three of the polypeptide sequences of 268, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 261 or the variable light chain sequence containing SEQ ID NO: 262; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:243, SEQ ID NO:245, SEQ ID NO:247 and SEQ ID NO:249, which correspond to SEQ ID NO: the heavy chain sequence of 241 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:242; and/or one of the polypeptide sequences of SEQ ID NO:263, SEQ ID NO:265, SEQ ID NO:267 and SEQ ID NO:269 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 261 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 262; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 241 or SEQ ID NO: 242 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 261 or SEQ ID NO: 262 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 244, SEQ ID NO: 246 and SEQ ID NO: 248 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 241 or the variable heavy chain sequence of SEQ ID NO: 242; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 264, SEQ ID NO: 266 and SEQ ID NO: 268 or three, which correspond to the light chain sequence of SEQ ID NO: 261 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO: 262; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 243, SEQ ID NO: 245, SEQ ID NO: 247 and SEQ ID NO: 249 Either, both, three or four, it corresponds to the heavy chain sequence of SEQ ID NO: 241 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 242; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 263, SEQ ID NO: 265, SEQ ID NO: 267 and SEQ ID NO: 269 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:261 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:262; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO: 242 variable heavy chain region; the variable light chain region of SEQ ID NO: 262; the complementarity determining region (SEQ ID NO: 244, SEQ ID NO: 246 and SEQ ID NO: 248) of the variable heavy chain region of SEQ ID NO: 242; and the variable of SEQ ID NO: 262 the complementarity determining region of the light chain region (SEQ ID NO: 264, SEQ ID NO: 266 and SEQ ID NO: 268); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO: 242 The variable heavy chain region; the variable light chain region of SEQ ID NO:262; the framework region of the variable heavy chain region of SEQ ID NO:242 (SEQ ID NO:243, SEQ ID NO:245, SEQ ID NO:247 and SEQ ID NO:249); and SEQ ID NO:262 The framework region of the variable light chain region (SEQ ID NO: 263, SEQ ID NO: 265, SEQ ID NO: 267 and SEQ ID NO: 269).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab7 comprising or consisting of SEQ ID NO: 241 and SEQ ID NO: 261; or an antibody comprising the CDRs of Ab7 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab7 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab7; or an antibody that binds to an epitope on HGF that is identical or overlapping with Ab7 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Ab7, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 242 and the variable light chain sequence of SEQ ID NO: 262 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO: 242 and/or SEQ ID NO: 262 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab7. In another embodiment of the present invention, anti-HGF antibodies such as Ab7 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab7 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab8

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：EVQLVESGGGLVQPGGSLRLSCAASGFTVSSNAISWVRQAPGKGLEWVGVIYVIGVTDYASSAQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVYDSGWNHFNLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:281)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: EVQLVESGGGLVQPGGSLRLSCAASGFTVSSNAISWVRQAPGKGLEWVGVIYVIGVTDYASSAQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVYDSGWNHFNLWGQGTLVTVSS (SEQ ID NO: 282).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab8相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 290).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：DIQMTQSPSTLSASVGDRVTITCQASQSISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQAYSVANVDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:301)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: DIQMTQSPSTLSASVGDRVTITCQASQSISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQAYSVANVDNAFGGGTKVEIKR (SEQ ID NO: 302).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab8 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 310).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO: 284, SEQ ID NO: 286 and SEQ ID NO: 288, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQ ID NO: 281 or it contains the variable heavy chain sequence of SEQ ID NO: 282; and/or it further contains SEQ ID NO: 304, SEQ ID NO: 306 and SEQ ID NO: One, two or three of the polypeptide sequences of 308, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 301 or the variable light chain sequence containing SEQ ID NO: 302; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:283, SEQ ID NO:285, SEQ ID NO:287 and SEQ ID NO:289, which correspond to SEQ ID NO: A heavy chain sequence of 281 or a framework region (FR or constant region) of a variable heavy chain sequence of SEQ ID NO:282; and/or one of the polypeptide sequences of SEQ ID NO:303, SEQ ID NO:305, SEQ ID NO:307 and SEQ ID NO:309 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 301 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 302; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 281 or SEQ ID NO: 282 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 301 or SEQ ID NO: 302 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 284, SEQ ID NO: 286 and SEQ ID NO: 288 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 281 or the variable heavy chain sequence of SEQ ID NO: 282; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 304, SEQ ID NO: 306 and SEQ ID NO: 308 or three, which correspond to the light chain sequence of SEQ ID NO: 301 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO: 302; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 283, SEQ ID NO: 285, SEQ ID NO: 287 and SEQ ID NO: 289 Either, both, three or four, it corresponds to the heavy chain sequence of SEQ ID NO: 281 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 282; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 303, SEQ ID NO: 305, SEQ ID NO: 307 and SEQ ID NO: 309 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:301 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:302; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: variable heavy chain region; the variable light chain region of SEQ ID NO:302; the complementarity determining region (SEQ ID NO:284, SEQ ID NO:286 and SEQ ID NO:288) of the variable heavy chain region of SEQ ID NO:282; and the variable of SEQ ID NO:302 the complementarity determining region of the light chain region (SEQ ID NO: 304, SEQ ID NO: 306 and SEQ ID NO: 308); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: Variable heavy chain region; variable light chain region of SEQ ID NO:302; framework region of variable heavy chain region of SEQ ID NO:282 (SEQ ID NO:283, SEQ ID NO:285, SEQ ID NO:287 and SEQ ID NO:289); and SEQ ID NO:302 The framework region of the variable light chain region (SEQ ID NO:303, SEQ ID NO:305, SEQ ID NO:307 and SEQ ID NO:309).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab8 comprising or consisting of SEQ ID NO: 281 and SEQ ID NO: 301; or an antibody comprising the CDRs of Ab8 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab8 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab8; or an antibody that binds to an epitope on HGF that is identical or overlapping with Ab8 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Concerning antibody Ab8, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 282 and the variable light chain sequence of SEQ ID NO: 302 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO: 282 and/or SEQ ID NO: 302, which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab8. In another embodiment of the present invention, anti-HGF antibodies such as Ab8 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab8 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab9

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSVEESGGRLVTPGTPLTLTCTVSGIDLNSNGMSWVRQAPGEGLEWIGASSIDGTTYYTNWAKGRFTISKTSSTTVDLKITSPTTEDTATYFCTRGEYAGVVGSNYFDLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:321)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSVEESGGRLVTPGTPLTLTCTVSGIDLNSNGMSWVRQAPGEGLEWIGASSIDGTTYYTNWAKGRFTISKTSSTTVDLKITSPTTEDTATYFCTRGEYAGVVGSNYFDLWGQGTLVTVSS (SEQ ID NO: 322).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab9相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 330).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：QVLTQTPPSVSAVVGGTVTINCQSSQRIYSNWLSWYQQKPGQTPKPLIYAASSLASGVPSRFKGSGSGTQFTLTISDLECDDAASYYCAGYYSGHIYSFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:341)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: QVLTQTPPSVSAVVGGTVTINCQSSQRIYSNWLSWYQQKPGQTPKPLIYAASSLASGVPSRFKGSGSGTQFTLTISDLECDDAASYYCAGYYSGHIYSFGGGTEVVVKR (SEQ ID NO: 342).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab9 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 350).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:324, SEQ ID NO:326 and SEQ ID NO:328, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO:321 or it contains the variable heavy chain sequence of SEQIDNO:322; And/it further contains SEQIDNO:344, SEQIDNO:346 and SEQIDNO: One, two or three of the polypeptide sequences of 348, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 341 or the variable light chain sequence containing SEQ ID NO: 342; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:323, SEQ ID NO:325, SEQ ID NO:327 and SEQ ID NO:329, which correspond to SEQ ID NO: the heavy chain sequence of 321 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:322; and/or one of the polypeptide sequences of SEQ ID NO:343, SEQ ID NO:345, SEQ ID NO:347 and SEQ ID NO:349 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 341 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 342; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 321 or SEQ ID NO: 322 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 341 or SEQ ID NO: 342 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 324, SEQ ID NO: 326 and SEQ ID NO: 328 or three, which correspond to the heavy chain sequence of SEQ ID NO: 321 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 322; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 344, SEQ ID NO: 346 and SEQ ID NO: 348 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 341 or the variable light chain sequence of SEQ ID NO: 342; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 323, SEQ ID NO: 325, SEQ ID NO: 327 and SEQ ID NO: 329 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO:321 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:322; Or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO:343, SEQ ID NO:345, SEQ ID NO:347 and SEQ ID NO:349 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:341 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:342; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: variable heavy chain region; the variable light chain region of SEQ ID NO:342; the complementarity determining region (SEQ ID NO:324, SEQ ID NO:326 and SEQ ID NO:328) of the variable heavy chain region of SEQ ID NO:322; and the variable of SEQ ID NO:342 the complementarity determining region of the light chain region (SEQ ID NO:344, SEQ ID NO:346 and SEQ ID NO:348); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: variable heavy chain region; variable light chain region of SEQ ID NO:342; framework region (SEQ ID NO:323, SEQ ID NO:325, SEQ ID NO:327 and SEQ ID NO:329) of the variable heavy chain region of SEQ ID NO:322; and SEQ ID NO:342 The framework region of the variable light chain region (SEQ ID NO: 343, SEQ ID NO: 345, SEQ ID NO: 347 and SEQ ID NO: 349).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab9 comprising or consisting of SEQ ID NO: 321 and SEQ ID NO: 341; or an antibody comprising the CDRs of Ab9 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab9 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab9; or an antibody that binds to an epitope on HGF that is identical or overlapping with Ab9 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Ab9, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 322 and the variable light chain sequence of SEQ ID NO: 342 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO: 322 and/or SEQ ID NO: 342 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab9. In another embodiment of the present invention, anti-HGF antibodies such as Ab9 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab9 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab10

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：EVQLVESGGGLVQPGGSLRLSCAASGFTVSSNGMSWVRQAPGKGLEWVGASSIDGTTYYTNSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGEYAGVVGSNYFDLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:361)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: EVQLVESGGGLVQPGGSLRLSCAASGFTVSSNGMSWVRQAPGKGLEWVGASSIDGTTYYTNSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGEYAGVVGSNYFDLWGQGTLVTVSS (SEQ ID NO: 362).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab10相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 370).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：DIQMTQSPSSVSASVGDRVTITCQSSQRIYSNWLSWYQQKPGKAPKLLIYAASSLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCAGYYSGHIYSFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:381)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: DIQMTQSPSSVSASVGDRVTITCQSSQRIYSNWLSWYQQKPGKAPKLLIYAASSLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCAGYYSGHIYSFGGGTKVEIKR (SEQ ID NO: 382).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab10 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 390).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:364, SEQ ID NO:366 and SEQ ID NO:368, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO:361 or it contains the variable heavy chain sequence of SEQIDNO:362; And/it further contains SEQIDNO:384, SEQIDNO:386 and SEQIDNO: One, two or three of the polypeptide sequences of 388, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 381 or the variable light chain sequence containing SEQ ID NO: 382; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:363, SEQ ID NO:365, SEQ ID NO:367 and SEQ ID NO:369, which correspond to SEQ ID NO: The heavy chain sequence of 361 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:362; , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 381 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 382; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 361 or SEQ ID NO: 362 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 381 or SEQ ID NO: 382 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 364, SEQ ID NO: 366 and SEQ ID NO: 368 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 361 or the variable heavy chain sequence of SEQ ID NO: 362; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 384, SEQ ID NO: 386 and SEQ ID NO: 388 or three, which correspond to the light chain sequence of SEQ ID NO: 381 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO: 382; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO:363, SEQ ID NO:365, SEQ ID NO:367 and SEQ ID NO:369 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO:361 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:362; Or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO:383, SEQ ID NO:385, SEQ ID NO:387 and SEQ ID NO:389 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:381 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:382; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: variable heavy chain region; the variable light chain region of SEQ ID NO:382; the complementarity determining region (SEQ ID NO:364, SEQ ID NO:366 and SEQ ID NO:368) of the variable heavy chain region of SEQ ID NO:362; and the variable of SEQ ID NO:382 the complementarity determining region of the light chain region (SEQ ID NO:384, SEQ ID NO:386 and SEQ ID NO:388); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: Variable heavy chain region; variable light chain region of SEQ ID NO:382; framework region of variable heavy chain region of SEQ ID NO:362 (SEQ ID NO:363, SEQ ID NO:365, SEQ ID NO:367 and SEQ ID NO:369); and SEQ ID NO:382 The framework region of the variable light chain region (SEQ ID NO:383, SEQ ID NO:385, SEQ ID NO:387 and SEQ ID NO:389).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab10 comprising or consisting of SEQ ID NO:361 and SEQ ID NO:381; or an antibody comprising the CDRs of Ab10 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab10 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab10; or an antibody that binds to an epitope identical or overlapping with that of Ab10 on HGF .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody AblO, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 362 and the variable light chain sequence of SEQ ID NO: 382 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO: 362 and/or SEQ ID NO: 382, which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of AblO. In another embodiment of the invention, anti-HGF antibodies such as Ab10 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab10 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab11

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSMEESGGRLVTPGTPLTLTCTVSGFSLSDYALSWVRQAPGKGLEWIGMISSGDNTYYASWAKGRFTISKASTTVDLKITSPTTEDTATYFCARDKDASSGGYLVLDLLDVPDGMDLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:401)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSMEESGGRLVTPGTPLTLTCTVSGFSLSDYALSWVRQAPGKGLEWIGMISSGDNTYYASWAKGRFTISKASTTVDLKITSPTTEDTATYFCARDKDASSGGYLVLDLLDVPDGMDLWGPGTLVTVSS (SEQ ID NO: 402).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab11相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 410).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：AVLTQTPSPVSAAVGGTVTIKCQSSQSVYNNNLLSWYQQKPGQPPKLLIWGASYLPSGVPDRFSGSGSGTQFTLTISGVQCDDAATYYCLGGYDGDADTYNTFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:421)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: AVLTQTPSPVSAAVGGTVTIKCQSSQSVYNNNLLSWYQQKPGQPPKLLIWGASYLPSGVPDRFSGSGSGTQFTLTISGVQCDDAATYYCLGGYDGDADTYNTFGGGTEVVVKR (SEQ ID NO: 422).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab11 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 430).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:404, SEQ ID NO:406 and SEQ ID NO:408, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO:401 or it contains the variable heavy chain sequence of SEQIDNO:402; and/or it further contains SEQIDNO:424, SEQIDNO:426 and SEQIDNO: One, two or three of the polypeptide sequences of 428, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 421 or the variable light chain sequence containing SEQ ID NO: 422; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:403, SEQ ID NO:405, SEQ ID NO:407 and SEQ ID NO:409, which correspond to SEQ ID NO: the heavy chain sequence of 401 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:402; and/or one of the polypeptide sequences of SEQ ID NO:423, SEQ ID NO:425, SEQ ID NO:427 and SEQ ID NO:429 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 421 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 422; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 401 or SEQ ID NO: 402 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 421 or SEQ ID NO: 422 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 404, SEQ ID NO: 406 and SEQ ID NO: 408 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 401 or the variable heavy chain sequence of SEQ ID NO: 402; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:424, SEQ ID NO:426 and SEQ ID NO:428 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 421 or the variable light chain sequence of SEQ ID NO: 422; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 403, SEQ ID NO: 405, SEQ ID NO: 407 and SEQ ID NO: 409 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO:401 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:402; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO:423, SEQ ID NO:425, SEQ ID NO:427 and SEQ ID NO:429 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:421 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:422; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: The variable heavy chain region; the variable light chain region of SEQ ID NO: 422; the complementarity determining region (SEQ ID NO: 404, SEQ ID NO: 406 and SEQ ID NO: 408) of the variable heavy chain region of SEQ ID NO: 402; and the variable of SEQ ID NO: 422 the complementarity determining region of the light chain region (SEQ ID NO:424, SEQ ID NO:426 and SEQ ID NO:428); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: variable heavy chain region; variable light chain region of SEQ ID NO:422; framework region (SEQ ID NO:403, SEQ ID NO:405, SEQ ID NO:407 and SEQ ID NO:409) of the variable heavy chain region of SEQ ID NO:402; and SEQ ID NO:422 The framework region of the variable light chain region (SEQ ID NO: 423, SEQ ID NO: 425, SEQ ID NO: 427 and SEQ ID NO: 429).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab11 comprising or consisting of SEQ ID NO: 401 and SEQ ID NO: 421; or an antibody comprising the CDRs of Ab11 and having at least one of the biological activities described herein or an antibody fragment; or an anti-HGF antibody that competes with Ab11 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab11; or an antibody that binds to an epitope identical or overlapping with Ab11 on HGF .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Concerning antibody Abl 1 , the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 402 and the variable light chain sequence of SEQ ID NO: 422 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:402 and/or SEQ ID NO:422 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl 1 . In another embodiment of the invention, anti-HGF antibodies such as Ab11 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab11 and one or more of the following fragments, variants Body, combination: FR, CDR, variable heavy chain and variable fragment: the variable heavy chain region of SEQIDNO:402; The variable light chain region of SEQIDNO:422; The framework region of the variable heavy chain region of SEQIDNO:402 ( SEQ ID NO:403, SEQ ID NO:405, SEQ ID NO:407 and SEQ ID NO:409); and the framework region of the variable light chain region of SEQ ID NO:422 (SEQ ID NO:423, SEQ ID NO:425, SEQ ID NO:427 and SEQ ID NO:429).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab11 comprising or consisting of SEQ ID NO: 401 and SEQ ID NO: 421; or an antibody comprising the CDRs of Ab11 and having at least one of the biological activities described herein or an antibody fragment; or an anti-HGF antibody that competes with Ab11 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab11; or an antibody that binds to an epitope identical or overlapping with Ab11 on HGF .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Concerning antibody Abl 1 , the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 402 and the variable light chain sequence of SEQ ID NO: 422 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:402 and/or SEQ ID NO:422 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl 1 . In another embodiment of the invention, anti-HGF antibodies such as Ab11 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab11 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab12

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSLEESGGRLVTPGGSLTLTCTVSGIDLSSNAISWVRQAPEKGLEWIAVIYVVGATDYASWAKGRFTISRTSTTVDLKMTSLTTEDTATYFCARVYDSGWNHFNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:441)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSLEESGGRLVTPGGSLTLTCTVSGIDLSSNAISWVRQAPEKGLEWIAVIYVVGATDYASWAKGRFTISRTSTTVDLKMTSLTTEDTATYFCARVYDSGWNHFNLWGPGTLVTVSS (SEQ ID NO: 442).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab12相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 450).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：AYDMTQTPASVEVAVGGTVTIKCQVSQSISSWLSWYQKKPGQRPKLLIYRASTLASGVSSRFKGSGSGTEFTLTISGVECADAATYYCQQAYSVSNVDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:461)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: AYDMTQTPASVEVAVGGTVTIKCQVSQSISSWLSWYQKKPGQRPKLLIYRASTLASGVSSRFKGSGSGTEFTLTISGVECADAATYYCQQAYSVSNVDNAFGGGTEVVVKR (SEQ ID NO: 462).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Abl2 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 470).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:444, SEQ ID NO:446 and SEQ ID NO:448, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQ ID NO: 441 or it contains the variable heavy chain sequence of SEQ ID NO: 442; and/or it further contains SEQ ID NO: 464, SEQ ID NO: 466 and SEQ ID NO: One, two or three of the polypeptide sequences of 468, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 461 or the variable light chain sequence containing SEQ ID NO: 462; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:443, SEQ ID NO:445, SEQ ID NO:447 and SEQ ID NO:449, which correspond to SEQ ID NO: the heavy chain sequence of 441 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:442; and/or one of the polypeptide sequences of SEQ ID NO:463, SEQ ID NO:465, SEQ ID NO:467 and SEQ ID NO:469 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 461 or the framework region of the variable light chain sequence of SEQ ID NO: 462; or these polypeptide sequences or are at least 80%, 90% or 95% identical thereto combination of sequences.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 441 or SEQ ID NO: 442 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 461 or SEQ ID NO: 462 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:444, SEQ ID NO:446 and SEQ ID NO:448 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 441 or the variable heavy chain sequence of SEQ ID NO: 442; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:464, SEQ ID NO:466 and SEQ ID NO:468 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 461 or the variable light chain sequence of SEQ ID NO: 462; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 443, SEQ ID NO: 445, SEQ ID NO: 447 and SEQ ID NO: 449 Either, both, three or four, it corresponds to the heavy chain sequence of SEQ ID NO: 441 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 442; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO:463, SEQ ID NO:465, SEQ ID NO:467 and SEQ ID NO:469 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:461 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:462; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: of SEQ ID NO:442 variable heavy chain region; the variable light chain region of SEQ ID NO:462; the complementarity determining region (SEQ ID NO:444, SEQ ID NO:446 and SEQ ID NO:448) of the variable heavy chain region of SEQ ID NO:442; and the variable of SEQ ID NO:462 The complementarity determining region of the light chain region (SEQ ID NO:464, SEQ ID NO:466 and SEQ ID NO:468) or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: of SEQ ID NO:442 variable heavy chain region; variable light chain region of SEQ ID NO:462; framework region (SEQ ID NO:443, SEQ ID NO:445, SEQ ID NO:447 and SEQ ID NO:449) of the variable heavy chain region of SEQ ID NO:442; and SEQ ID NO:462 The framework region of the variable light chain region (SEQ ID NO: 463, SEQ ID NO: 465, SEQ ID NO: 467 and SEQ ID NO: 469).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab12 comprising or consisting of SEQ ID NO:441 and SEQ ID NO:461; or an antibody comprising the CDRs of Ab12 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Abl2 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Abl2; or an antibody that binds to an epitope identical or overlapping with Abl2 on HGF .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Abl2, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 442 and the variable light chain sequence of SEQ ID NO: 462 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:442 and/or SEQ ID NO:462 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl2. In another embodiment of the invention, anti-HGF antibodies such as Abl2 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab12 and the FR, CDR, variable heavy chain as described above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab13

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：EVQLVESGGGLVQPGGSLRLSCAASGFTVSSNAISWVRQAPGKGLEWVAVIYVVGATDYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVYDSGWNHFNLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:481)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: EVQLVESGGGLVQPGGSLRLSCAASGFTVSSNAISWVRQAPGKGLEWVAVIYVVGATDYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVYDSGWNHFNLWGQGTLVTVSS (SEQ ID NO: 482).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab13相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 490).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：DYQMTQSPSTLSASVGDRVTITCQVSQSISSWLSWYQQKPGKAPKLLIYRASTLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQAYSVSNVDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:501)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: DYQMTQSPSTLSASVGDRVTITCQVSQSISSWLSWYQQKPGKAPKLLIYRASTLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQAYSVSNVDNAFGGGTKVEIKR (SEQ ID NO: 502).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind to the same epitope as Ab13 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 510).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:484, SEQ ID NO:486 and SEQ ID NO:488, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO:481 or it contains the variable heavy chain sequence of SEQIDNO:482; and/or it further contains SEQIDNO:504, SEQIDNO:506 and SEQIDNO: One, two or three of the polypeptide sequences of 508, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 501 or the variable light chain sequence containing SEQ ID NO: 502; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:483, SEQ ID NO:485, SEQ ID NO:487 and SEQ ID NO:489, which correspond to SEQ ID NO: the heavy chain sequence of 481 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:482; and/or one of the polypeptide sequences of SEQ ID NO:503, SEQ ID NO:505, SEQ ID NO:507 and SEQ ID NO:509 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 501 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 502; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 481 or SEQ ID NO: 482 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 501 or SEQ ID NO: 502 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:484, SEQ ID NO:486 and SEQ ID NO:488 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 481 or the variable heavy chain sequence of SEQ ID NO: 482; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:504, SEQ ID NO:506 and SEQ ID NO:508 or three, which correspond to the light chain sequence of SEQ ID NO:501 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO:502; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 483, SEQ ID NO: 485, SEQ ID NO: 487 and SEQ ID NO: 489 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO:481 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:482; Or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO:503, SEQ ID NO:505, SEQ ID NO:507, and SEQ ID NO:509 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:501 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:502; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: of SEQ ID NO:482 variable heavy chain region; the variable light chain region of SEQ ID NO:502; the complementarity determining region (SEQ ID NO:484, SEQ ID NO:486 and SEQ ID NO:488) of the variable heavy chain region of SEQ ID NO:482; and the variable of SEQ ID NO:502 the complementarity determining region of the light chain region (SEQ ID NO:504, SEQ ID NO:506 and SEQ ID NO:508); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: of SEQ ID NO:482 Variable heavy chain region; variable light chain region of SEQ ID NO:502; framework region of variable heavy chain region of SEQ ID NO:482 (SEQ ID NO:483, SEQ ID NO:485, SEQ ID NO:487 and SEQ ID NO:489); and SEQ ID NO:502 The framework region of the variable light chain region (SEQ ID NO:503, SEQ ID NO:505, SEQ ID NO:507 and SEQ ID NO:509).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab13 comprising or consisting of SEQ ID NO:481 and SEQ ID NO:501; or an antibody comprising the CDRs of Ab13 and having at least one of the biological activities described herein or an antibody fragment; or an anti-HGF antibody that competes with Abl3 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Abl3; or an antibody that binds to an epitope identical or overlapping with Abl3 on HGF .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Abl3, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 482 and the variable light chain sequence of SEQ ID NO: 502 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:482 and/or SEQ ID NO:502 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl3. In another embodiment of the invention, anti-HGF antibodies such as Abl3 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab13 and the FR, CDR, variable heavy chain as described above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab14

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSVEESGGRLVTPGTPLTLTCTVSGFSLSNYAMTWVRQAPGKGLEWIGVISFGGNTYYANWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARWDAENNEILNLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:521)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSVEESGGRLVTPGTPLTLTCTVSGFSLSNYAMTWVRQAPGKGLEWIGVISFGGNTYYANWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARWDAENNEILNLWGQGTLVTVSS (SEQ ID NO: 522).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab14相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 530).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：AYDMTQTPASVEVAVGGTVTIKCQASESIESYLAWYQQKSGQPPKLLIYRASTLASGVSSRFKGSGSGTQFTLTISGVECADAATYYCQQGDAWSNVDNVFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:541)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: AYDMTQTPASVEVAVGGTVTIKCQASESIESYLAWYQQKSGQPPKLLIYRASTLASGVSSRFKGSGSGTQFTLTISGVECADAATYYCQQGDAWSNVDNVFGGGTEVVVKR (SEQ ID NO: 542).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Abl4 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 550).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:524, SEQ ID NO:526 and SEQ ID NO:528, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO:521 or it contains the variable heavy chain sequence of SEQIDNO:522; and/or it further contains SEQIDNO:544, SEQIDNO:546 and SEQIDNO: One, two or three of the polypeptide sequences of 548, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 541 or the variable light chain sequence containing SEQ ID NO: 542; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:523, SEQ ID NO:525, SEQ ID NO:527 and SEQ ID NO:529, which correspond to SEQ ID NO: the heavy chain sequence of 521 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:522; and/or one of the polypeptide sequences of SEQ ID NO:543, SEQ ID NO:545, SEQ ID NO:547 and SEQ ID NO:549 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 541 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 542; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO:521 or SEQ ID NO:522 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 541 or SEQ ID NO: 542 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:524, SEQ ID NO:526 and SEQ ID NO:528 or three, which correspond to the heavy chain sequence of SEQ ID NO:521 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO:522; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:544, SEQ ID NO:546 and SEQ ID NO:548 or three, which correspond to the light chain sequence of SEQ ID NO:541 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO:542; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO:523, SEQ ID NO:525, SEQ ID NO:527 and SEQ ID NO:529 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO:521 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:522; Or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO:543, SEQ ID NO:545, SEQ ID NO:547 and SEQ ID NO:549 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:541 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:542; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO:522 The variable heavy chain region; the variable light chain region of SEQ ID NO:542; the complementarity determining region (SEQ ID NO:524, SEQ ID NO:526 and SEQ ID NO:528) of the variable heavy chain region of SEQ ID NO:522; and the variable of SEQ ID NO:542 the complementarity determining region of the light chain region (SEQ ID NO:544, SEQ ID NO:546 and SEQ ID NO:548); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO:522 Variable heavy chain region; variable light chain region of SEQ ID NO:542; framework region of variable heavy chain region of SEQ ID NO:522 (SEQ ID NO:523, SEQ ID NO:525, SEQ ID NO:527 and SEQ ID NO:529); and SEQ ID NO:542 The framework region of the variable light chain region (SEQ ID NO:543, SEQ ID NO:545, SEQ ID NO:547 and SEQ ID NO:549).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab14 comprising or consisting of SEQ ID NO:521 and SEQ ID NO:541; or an antibody comprising the CDRs of Ab14 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Abl4 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Abl4; or an antibody that binds to an epitope identical or overlapping with Abl4 on HGF .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Abl4, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO:522 and the variable light chain sequence of SEQ ID NO:542 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:522 and/or SEQ ID NO:542 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl4. In another embodiment of the present invention, anti-HGF antibodies such as Abl4 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab14 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab15

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSVEESGGRLVTPGTPLTLTCTVSGFSLSNYAMTWVRQAPGKGLEWIGVISFGGNTYYANWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARWDAENNEILNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:561)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSVEESGGRLVTPGTPLTLTCTVSGFSLSNYAMTWVRQAPGKGLEWIGVISFGGNTYYANWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARWDAENNEILNLWGPGTLVTVSS (SEQ ID NO: 562).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab15相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 570).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：AYDMTQTPASVEVAVGGTVTIKCQASESISSYLAWYQQKSGQPPKLLIYRASTLASGVSSRFKGSGSGTQFTLTISGVECADAATYYCQQGDAWSNVDNVFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:581)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: AYDMTQTPASVEVAVGGTVTIKCQASESISSYLAWYQQKSGQPPKLLIYRASTLASGVSSRFKGSGSGTQFTLTISGVECADAATYYCQQGDAWSNVDNVFGGGTEVVVKR (SEQ ID NO: 582).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab15 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 590).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:564, SEQ ID NO:566 and SEQ ID NO:568, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQ ID NO: 561 or it contains the variable heavy chain sequence of SEQ ID NO: 562; and/or it further contains SEQ ID NO: 584, SEQ ID NO: 586 and SEQ ID NO: One, two or three of the polypeptide sequences of 588, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 581 or the variable light chain sequence containing SEQ ID NO: 582; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:563, SEQ ID NO:565, SEQ ID NO:567 and SEQ ID NO:569, which correspond to SEQ ID NO: the heavy chain sequence of 561 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:562; and/or one of the polypeptide sequences of SEQ ID NO:583, SEQ ID NO:585, SEQ ID NO:587 and SEQ ID NO:589 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 581 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 582; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO:561 or SEQ ID NO:562 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 581 or SEQ ID NO: 582 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:564, SEQ ID NO:566 and SEQ ID NO:568 or three, which correspond to the heavy chain sequence of SEQ ID NO:561 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO:562; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:584, SEQ ID NO:586 and SEQ ID NO:588 or three, which correspond to the light chain sequence of SEQ ID NO:581 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO:582; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO:563, SEQ ID NO:565, SEQ ID NO:567 and SEQ ID NO:569 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO:561 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:562; Or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO:583, SEQ ID NO:585, SEQ ID NO:587 and SEQ ID NO:589 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:581 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:582; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO:562 variable heavy chain region; the variable light chain region of SEQ ID NO:582; the complementarity determining region (SEQ ID NO:564, SEQ ID NO:566 and SEQ ID NO:568) of the variable heavy chain region of SEQ ID NO:562; and the variable of SEQ ID NO:582 the complementarity determining region of the light chain region (SEQ ID NO:584, SEQ ID NO:586 and SEQ ID NO:588); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO:562 Variable heavy chain region; variable light chain region of SEQ ID NO:582; framework region of variable heavy chain region of SEQ ID NO:562 (SEQ ID NO:563, SEQ ID NO:565, SEQ ID NO:567 and SEQ ID NO:569); and SEQ ID NO:582 The framework region of the variable light chain region (SEQ ID NO:583, SEQ ID NO:585, SEQ ID NO:587 and SEQ ID NO:589).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab15 comprising or consisting of SEQ ID NO:561 and SEQ ID NO:581; or an antibody comprising the CDRs of Ab15 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Abl5 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Abl5; or an antibody that binds to an epitope identical or overlapping with Abl5 on HGF .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Concerning antibody Abl5, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 562 and the variable light chain sequence of SEQ ID NO: 582 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:562 and/or SEQ ID NO:582 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl5. In another embodiment of the invention, anti-HGF antibodies such as Abl5 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab15 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab16

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSVEESGGRLVTPGTPLTLTCTVSGIDLSNYAMGWVRQAPGKGLEYIGMIGVNGRAWYATWAKGRFTISKTSPTVDLKITSPTTEDTATYFCARLIDERSTYSYVFDLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:601)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSVEESGGRLVTPGTPLTLTCTVSGIDLSNYAMGWVRQAPGKGLEYIGMIGVNGRAWYATWAKGRFTISKTSPTVDLKITSPTTEDTATYFCARLIDERSTYSYVFDLWGQGTLVTVSS (SEQ ID NO: 602).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab16相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 610).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：QVLTQTPSPVSAAVGGTVTINCQGSQSLYNNNAFSWYQQKPGQPPKLLIYDASTLASGVPSRFKGSGSGTQFTLTISGVQCADAATYYCQGEFSCGDVDCIAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:621)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: QVLTQTPSPVSAAVGGTVTINCQGSQSLYNNNAFSWYQQKPGQPPKLLIYDASTLASGVPSRFKGSGSGTQFTLTISGVQCADAATYYCQGEFSCGDVDCIAFGGGTEVVVKR (SEQ ID NO: 622).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab16 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 630).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:604, SEQ ID NO:606 and SEQ ID NO:608, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO:601 or it contains the variable heavy chain sequence of SEQIDNO:602; and/or it further contains SEQIDNO:624, SEQIDNO:626 and SEQIDNO: One, two or three of the polypeptide sequences of 628, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 621 or the variable light chain sequence containing SEQ ID NO: 622; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:603, SEQ ID NO:605, SEQ ID NO:607 and SEQ ID NO:609, which correspond to SEQ ID NO: The heavy chain sequence of 601 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:602; and/or one of the polypeptide sequences of SEQ ID NO:623, SEQ ID NO:625, SEQ ID NO:627 and SEQ ID NO:629 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 621 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 622; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 601 or SEQ ID NO: 602 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 621 or SEQ ID NO: 622 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:604, SEQ ID NO:606 and SEQ ID NO:608 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 601 or the variable heavy chain sequence of SEQ ID NO: 602; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:624, SEQ ID NO:626 and SEQ ID NO:628 or three, which correspond to the light chain sequence of SEQ ID NO:621 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO:622; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO:603, SEQ ID NO:605, SEQ ID NO:607 and SEQ ID NO:609 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO:601 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:602; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO:623, SEQ ID NO:625, SEQ ID NO:627 and SEQ ID NO:629 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:621 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:622; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: variable heavy chain region; the variable light chain region of SEQ ID NO:622; the complementarity determining region (SEQ ID NO:604, SEQ ID NO:606 and SEQ ID NO:608) of the variable heavy chain region of SEQ ID NO:602; and the variable of SEQ ID NO:622 the complementarity determining region of the light chain region (SEQ ID NO:624, SEQ ID NO:626 and SEQ ID NO:628); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: variable heavy chain region; variable light chain region of SEQ ID NO:622; framework region (SEQ ID NO:603, SEQ ID NO:605, SEQ ID NO:607 and SEQ ID NO:609) of the variable heavy chain region of SEQ ID NO:602; and SEQ ID NO:622 The framework region of the variable light chain region (SEQ ID NO:623, SEQ ID NO:625, SEQ ID NO:627 and SEQ ID NO:629).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab16 comprising or consisting of SEQ ID NO:601 and SEQ ID NO:621; or an antibody comprising the CDRs of Ab16 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab16 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab16; or an antibody that binds to an epitope identical or overlapping with Ab16 on HGF .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Concerning antibody Abl6, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 602 and the variable light chain sequence of SEQ ID NO: 622 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:602 and/or SEQ ID NO:622 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl6. In another embodiment of the invention, anti-HGF antibodies such as Ab16 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab16 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab17

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSVEESGGRLVPPGTPLTLTCTVSGIDLSSYAMGWVRQAPGKGLEYIGMIDVSGSTYYADWAKGRLTISKTPTTVDLEITSPTTEDTATYFCARLIDERSTYSYAFDLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:641)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSVEESGGRLVPPPGTPLTLTCTVSGIDLSSYAMGWVRQAPGKGLEYIGMIDVSGSTYYADWAKGRLTISKTPTTVDLEITSPTTEDTATYFCARLIDERSTYSYAFDLWGQGTLVTVSS (SEQ ID NO: 642).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab17相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 650).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：QVLTQTPSPVSAAVGGTVTINCQASQSFYNNGAFSWYQQKPGQPPKLLIYDASTLASGVPSRFKGSGSGTQFTLTISGVQCGDAATYYCQGEFSCGSADCVAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:661)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: QVLTQTPSPVSAAVGGTVTINCQASQSFYNNGAFSWYQQKPGQPPKLLIYDASTLASGVPSRFKGSGSGTQFTLTISGVQCGDAATYYCQGEFSCGSADCVAFGGGTEVVVKR (SEQ ID NO: 662).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Abl7 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 670).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:644, SEQ ID NO:646 and SEQ ID NO:648, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQ ID NO: 641 or it contains the variable heavy chain sequence of SEQ ID NO: 642; and/or it further contains SEQ ID NO: 664, SEQ ID NO: 666 and SEQ ID NO: One, two or three of the polypeptide sequences of 668, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 661 or the variable light chain sequence containing SEQ ID NO: 662; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:643, SEQ ID NO:645, SEQ ID NO:647 and SEQ ID NO:649, which correspond to SEQ ID NO: the heavy chain sequence of 641 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:642; and/or one of the polypeptide sequences of SEQ ID NO:663, SEQ ID NO:665, SEQ ID NO:667 and SEQ ID NO:669 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 661 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 662; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 641 or SEQ ID NO: 642 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 661 or SEQ ID NO: 662 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:644, SEQ ID NO:646 and SEQ ID NO:648 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 641 or the variable heavy chain sequence of SEQ ID NO: 642; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:664, SEQ ID NO:666 and SEQ ID NO:668 or three, which correspond to the light chain sequence of SEQ ID NO: 661 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO: 662; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO:643, SEQ ID NO:645, SEQ ID NO:647 and SEQ ID NO:649 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO:641 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:642; Or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO:663, SEQ ID NO:665, SEQ ID NO:667 and SEQ ID NO:669 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:661 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:662; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: of SEQ ID NO:642 variable heavy chain region; the variable light chain region of SEQ ID NO:662; the complementarity determining region (SEQ ID NO:644, SEQ ID NO:646 and SEQ ID NO:648) of the variable heavy chain region of SEQ ID NO:642; and the variable of SEQ ID NO:662 the complementarity determining region of the light chain region (SEQ ID NO:664, SEQ ID NO:666 and SEQ ID NO:668); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: of SEQ ID NO:642 Variable heavy chain region; variable light chain region of SEQ ID NO:662; framework region of variable heavy chain region of SEQ ID NO:642 (SEQ ID NO:643, SEQ ID NO:645, SEQ ID NO:647 and SEQ ID NO:649); and SEQ ID NO:662 The framework region of the variable light chain region (SEQ ID NO: 663, SEQ ID NO: 665, SEQ ID NO: 667 and SEQ ID NO: 669).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab17 comprising or consisting of SEQ ID NO:641 and SEQ ID NO:661; or an antibody comprising the CDRs of Ab17 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Abl7 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Abl7; or an antibody that binds to an epitope identical or overlapping with Abl7 on HGF .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Abl7, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 642 and the variable light chain sequence of SEQ ID NO: 662 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:642 and/or SEQ ID NO:662 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl7. In another embodiment of the invention, anti-HGF antibodies such as Abl7 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab17 and the FR, CDR, variable heavy chain as described above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab18

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSLEESGGRLVTPGTPLTLTCTASGFSLSSYDMSWVRQAPGKGLEWIGIIYAGSASTWFASWVKGRFTISKTSTTVDLKMTSLTTEDTATYFCARVGYSGYGYDDNLDMWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:681)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSLEESGGRLVTPGTPLTLTCTASGFSLSSYDMSWVRQAPGKGLEWIGIIYAGSASTWFASWVKGRFTISKTSTTVDLKMTSLTTEDTATYFCARVGYSGYDDNLDMWGQGTLVTVSS (SEQ ID NO: 682).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab18相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 690).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：AYDMTQTPASVEVAVGGTVTIKCQASQSISTALAWYQQKPGQRPKLLIYDASKLASGVSSRFKGSGSGAQFTLTISGVECADAATYYCHQGYSSSNVDNTFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:701)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: AYDMTQTPASVEVAVGGTVTIKCQASQSISTALAWYQQKPGQRPKLLIYDASKLASGVSSRFKGSGSGAQFTLTISGVECADAATYYCHQGYSSSNVDNTFGGGTEVVVKR (SEQ ID NO: 702).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab18 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 710).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:684, SEQ ID NO:686 and SEQ ID NO:688, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO:681 or it contains the variable heavy chain sequence of SEQIDNO:682; and/or it further contains SEQIDNO:704, SEQIDNO:706 and SEQIDNO: One, two or three of the polypeptide sequences of 708, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 701 or the variable light chain sequences containing SEQ ID NO: 702; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:683, SEQ ID NO:685, SEQ ID NO:687 and SEQ ID NO:689, which correspond to SEQ ID NO: the heavy chain sequence of 681 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:682; and/or one of the polypeptide sequences of SEQ ID NO:703, SEQ ID NO:705, SEQ ID NO:707 and SEQ ID NO:709 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 701 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 702; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 681 or SEQ ID NO: 682 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 701 or SEQ ID NO: 702 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:684, SEQ ID NO:686 and SEQ ID NO:688 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 681 or the variable heavy chain sequence of SEQ ID NO: 682; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 704, SEQ ID NO: 706 and SEQ ID NO: 708 or three, which correspond to the light chain sequence of SEQ ID NO:701 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO:702; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO:683, SEQ ID NO:685, SEQ ID NO:687 and SEQ ID NO:689 Either, two, three or four, it corresponds to the heavy chain sequence of SEQIDNO:681 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:682; Or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO:703, SEQ ID NO:705, SEQ ID NO:707 and SEQ ID NO:709 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:701 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:702; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: The variable heavy chain region; the variable light chain region of SEQ ID NO: 702; the complementarity determining region (SEQ ID NO: 684, SEQ ID NO: 686 and SEQ ID NO: 688) of the variable heavy chain region of SEQ ID NO: 682; and the variable of SEQ ID NO: 702 the complementarity determining region of the light chain region (SEQ ID NO: 704, SEQ ID NO: 706 and SEQ ID NO: 708); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: Variable heavy chain region; variable light chain region of SEQ ID NO:702; framework region of variable heavy chain region of SEQ ID NO:682 (SEQ ID NO:683, SEQ ID NO:685, SEQ ID NO:687 and SEQ ID NO:689); and SEQ ID NO:702 The framework region of the variable light chain region (SEQ ID NO: 703, SEQ ID NO: 705, SEQ ID NO: 707 and SEQ ID NO: 709).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab18 comprising or consisting of SEQ ID NO:681 and SEQ ID NO:701; or an antibody comprising the CDRs of Ab18 and having at least one of the biological activities described herein or an antibody fragment; or an anti-HGF antibody that competes with Abl8 for binding to HGF, preferably an antibody that contains a sequence at least 90% or 95% identical to the sequence of Abl8; or an antibody that binds to an epitope identical or overlapping with Abl8 on HGF .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Abl8, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 682 and the variable light chain sequence of SEQ ID NO: 702 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:682 and/or SEQ ID NO:702 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl8. In another embodiment of the invention, anti-HGF antibodies such as Abl8 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab18 and the FR, CDR, variable heavy chain as described above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab19

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSVEESGGRLVTPGTPLTLTCTASGFSLSNYWMGWVRQAPGEGLEWIGTISYDGNTYYASWAKGRFTISRTSTTVDLKMTSLTTEDTAIYFCATVNYPDYSTGAFNIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:721)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSVEESGGRLVTPGTPLTLTTCASGFSLSNYWMGWVRQAPGEGLEWIGTISYDGNTYYASWAKGRFTISRTSTTVDLKMTSLTTEDTAIYFCATVNYPDYSTGAFNIWGPGTLVTVSS (SEQ ID NO: 722).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab19相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 730).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：DVVMTQTPASVSEPVGGTVTIKCQASQSIDNYLAWYQQKPGQRPRLLIYYTSTLASGVPSRFKGSGSGTEYTLTISDLECADAATYYCQFTAYYSTYIGAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:741)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: DVVMTQTPASVSEPVGGTVTIKCQASQSIDNYLAWYQQKPGQRPRLLIYYTSTLASGVPSRFKGSGSGTEYTLTISDLECADATYYCQFTAYYSTYIGAFGGGTEVVVKR (SEQ ID NO: 742).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab19 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 750).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:724, SEQ ID NO:726 and SEQ ID NO:728, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO:721 or it contains the variable heavy chain sequence of SEQIDNO:722; And/it further contains SEQIDNO:744, SEQIDNO:746 and SEQIDNO: One, two or three of the polypeptide sequences of 748, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 741 or the variable light chain sequence containing SEQ ID NO: 742; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:723, SEQ ID NO:725, SEQ ID NO:727 and SEQ ID NO:729, which correspond to SEQ ID NO: the heavy chain sequence of 721 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:722; and/or one of the polypeptide sequences of SEQ ID NO:743, SEQ ID NO:745, SEQ ID NO:747 and SEQ ID NO:749 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 741 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 742; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 721 or SEQ ID NO: 722 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 741 or SEQ ID NO: 742 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:724, SEQ ID NO:726 and SEQ ID NO:728 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 721 or the variable heavy chain sequence of SEQ ID NO: 722; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:744, SEQ ID NO:746 and SEQ ID NO:748 or three, which correspond to the light chain sequence of SEQ ID NO:741 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO:742; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 723, SEQ ID NO: 725, SEQ ID NO: 727 and SEQ ID NO: 729 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO:721 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:722; Or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO:743, SEQ ID NO:745, SEQ ID NO:747 and SEQ ID NO:749 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:741 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:742; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO:722 variable heavy chain region; the variable light chain region of SEQ ID NO:742; the complementarity determining region (SEQ ID NO:724, SEQ ID NO:726 and SEQ ID NO:728) of the variable heavy chain region of SEQ ID NO:722; and the variable of SEQ ID NO:742 the complementarity determining region of the light chain region (SEQ ID NO: 744, SEQ ID NO: 746 and SEQ ID NO: 748); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO:722 variable heavy chain region; variable light chain region of SEQ ID NO:742; framework region (SEQ ID NO:723, SEQ ID NO:725, SEQ ID NO:727 and SEQ ID NO:729) of the variable heavy chain region of SEQ ID NO:722; and SEQ ID NO:742 The framework region of the variable light chain region (SEQ ID NO: 743, SEQ ID NO: 745, SEQ ID NO: 747 and SEQ ID NO: 749).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab19 comprising or consisting of SEQ ID NO:721 and SEQ ID NO:741; or an antibody comprising the CDRs of Ab19 and having at least one of the biological activities described herein or an antibody fragment; or an anti-HGF antibody that competes with Ab19 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab19; or an antibody that binds to an epitope that is identical or overlapping with Ab19 on HGF .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Abl9, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 722 and the variable light chain sequence of SEQ ID NO: 742 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:722 and/or SEQ ID NO:742 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl9. In another embodiment of the invention, anti-HGF antibodies such as Ab19 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab19 and the FR, CDR, variable heavy chain as described above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab20

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：EVQLVESGGGLVQPGGSLRLSCAASGFTVSNYWMGWVRQAPGKGLEWIGTISYDGNTYYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCATVNYPDYSTGAFNIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:761)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: EVQLVESGGGLVQPGGSLRLSCAASGFTVSNYWMGWVRQAPGKGLEWIGTISYDGNTYYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCATVNYPDYSTGAFNIWGQGTLVTVSS (SEQ ID NO: 762).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab20相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 770).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：DIQMTQSPSSLSASVGDRVTITCQASQSIDNYLAWYQQKPGKVPKLLIYYTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQFTAYYSTYIGAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:781)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: DIQMTQSPSSLSASVGDRVTITCQASQSIDNYLAWYQQKPGKVPKLLIYYTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQFTAYYSTYIGAFGGGTKVEIKR (SEQ ID NO: 782).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab20 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 790).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:764, SEQ ID NO:766 and SEQ ID NO:768, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO:761 or it contains the variable heavy chain sequence of SEQIDNO:762; and/or it further contains SEQIDNO:784, SEQIDNO:786 and SEQIDNO: One, two or three of the polypeptide sequences of 788, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 781 or the variable light chain sequence containing SEQ ID NO: 782; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:763, SEQ ID NO:765, SEQ ID NO:767 and SEQ ID NO:769, which correspond to SEQ ID NO: the heavy chain sequence of 761 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:762; and/or one of the polypeptide sequences of SEQ ID NO:783, SEQ ID NO:785, SEQ ID NO:787 and SEQ ID NO:789 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 781 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 782; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 761 or SEQ ID NO: 762 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 781 or SEQ ID NO: 782 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:764, SEQ ID NO:766 and SEQ ID NO:768 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 761 or the variable heavy chain sequence of SEQ ID NO: 762; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 784, SEQ ID NO: 786 and SEQ ID NO: 788 or three, which correspond to the light chain sequence of SEQ ID NO:781 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO:782; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 763, SEQ ID NO: 765, SEQ ID NO: 767 and SEQ ID NO: 769 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO: 761 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO: 762; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 783, SEQ ID NO: 785, SEQ ID NO: 787 and SEQ ID NO: 789 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:781 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:782; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO:762 The variable heavy chain region; the variable light chain region of SEQ ID NO: 782; the complementarity determining region (SEQ ID NO: 764, SEQ ID NO: 766 and SEQ ID NO: 768) of the variable heavy chain region of SEQ ID NO: 762; and the variable of SEQ ID NO: 782 the complementarity determining region of the light chain region (SEQ ID NO: 784, SEQ ID NO: 786 and SEQ ID NO: 788); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO:762 Variable heavy chain region; variable light chain region of SEQ ID NO:782; framework region of variable heavy chain region of SEQ ID NO:762 (SEQ ID NO:763, SEQ ID NO:765, SEQ ID NO:767 and SEQ ID NO:769); and SEQ ID NO:782 The framework regions of the variable light chain region (SEQ ID NO: 783, SEQ ID NO: 785, SEQ ID NO: 787 and SEQ ID NO: 789).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab20 comprising or consisting of SEQ ID NO: 761 and SEQ ID NO: 781; or an antibody comprising the CDRs of Ab20 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab20 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab20; or an antibody that binds to an epitope on HGF that is identical or overlaps with Ab20 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Ab20, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 762 and the variable light chain sequence of SEQ ID NO: 782 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:762 and/or SEQ ID NO:782 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab20. In another embodiment of the present invention, anti-HGF antibodies such as Ab20 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab20 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab21

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSVEESGGRLVTPGTPLTLTCTVSGFSLSTYYMSWVRQAPGKGLEWIGIIYVSGITDYARWAKGRFTISKTSTTVDLKMTSLTTEDTATYFCARHIDSSGWDGLGIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:801)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSVEESGGRLVTPGTPLTLTCTVSGFSLSTYYMSWVRQAPGKGLEWIGIIYVSGITDYARWAKGRFTISKTSTTVDLKMTSLTTEDTATYFCARHIDSSGWDGLGIWGQGTLVTVSS (SEQ ID NO: 802).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab21相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 810).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：AYDMTQTPASVEVAVGGTVTIKCQASESISSYLNWYQQKLGQPPKLLIYRASTLTSGVSSRFKGSGSGTEYTLTISDLECADAATYYCQQTYGYSDTDNSFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:821)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: AYDMTQTPASVEVAVGGTVTIKCQASESISSYLNWYQQKLGQPPKLLIYRASTLTSGVSSRFKGSGSGTEYTLTISDLECADATYYCQQTYGYSDTDNSFGGGTEVVVKR (SEQ ID NO: 822).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab21 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 830).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:804, SEQ ID NO:806 and SEQ ID NO:808, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO:801 or it contains the variable heavy chain sequence of SEQIDNO:802; and/or it further contains SEQIDNO:824, SEQIDNO:826 and SEQIDNO: One, two or three of the polypeptide sequences of 828, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 821 or the variable light chain sequence containing SEQ ID NO: 822; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:803, SEQ ID NO:805, SEQ ID NO:807 and SEQ ID NO:809, which correspond to SEQ ID NO: the heavy chain sequence of 801 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:802; and/or one of the polypeptide sequences of SEQ ID NO:823, SEQ ID NO:825, SEQ ID NO:827 and SEQ ID NO:829 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 821 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 822; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 801 or SEQ ID NO: 802 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 821 or SEQ ID NO: 822 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:804, SEQ ID NO:806 and SEQ ID NO:808 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 801 or the variable heavy chain sequence of SEQ ID NO: 802; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:824, SEQ ID NO:826 and SEQ ID NO:828 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 821 or the variable light chain sequence of SEQ ID NO: 822; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO:803, SEQ ID NO:805, SEQ ID NO:807 and SEQ ID NO:809 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO:801 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:802; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 823, SEQ ID NO: 825, SEQ ID NO: 827 and SEQ ID NO: 829 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:821 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:822; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: variable heavy chain region; the variable light chain region of SEQ ID NO:822; the complementarity determining region (SEQ ID NO:804, SEQ ID NO:806 and SEQ ID NO:808) of the variable heavy chain region of SEQ ID NO:802; and the variable of SEQ ID NO:822 the complementarity determining region of the light chain region (SEQ ID NO:824, SEQ ID NO:826 and SEQ ID NO:828); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: Variable heavy chain region; the variable light chain region of SEQ ID NO:822; the framework region (SEQ ID NO:803, SEQ ID NO:805, SEQ ID NO:807 and SEQ ID NO:809) of the variable heavy chain region of SEQ ID NO:802; Variable light chain regions and framework regions (SEQ ID NO:823, SEQ ID NO:825, SEQ ID NO:827 and SEQ ID NO:829).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab21 comprising or consisting of SEQ ID NO: 801 and SEQ ID NO: 821; or an antibody comprising the CDRs of Ab21 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab21 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab21; or an antibody that binds to an epitope on HGF that is identical or overlaps with Ab21 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Ab21, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 802 and the variable light chain sequence of SEQ ID NO: 822 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:802 and/or SEQ ID NO:822 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab21. In another embodiment of the present invention, anti-HGF antibodies such as Ab21 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab21 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab23

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSLEESGGRLVTPGTPLTLTCTASGFTIGRYYMSWVRQAPGKGLEWIGIIYTHGVNPDYASWAKGRFTISRPSTTVDLKITSPTTEDTATYFCARVGGFNDYSDIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:841)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSLEESGGRLVTPGTPLTLTCTASGFTIGRYYMSWVRQAPGKGLEWIGIIYTHGVNPDYASWAKGRFTISRPSTTVDLKITSPTTEDTATYFCARVGGFNDYSDIWGPGTLVTVSS (SEQ ID NO: 842).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab23相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 850).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：AYDMTQTPASVEVAVGGTVTIKCQASESISTYLAWYQQKPGQPPKLLIYRASTLASGVSSRFKGSGSGTQFTLTISGVECADAATYYCQQGYSYSNVDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:861)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: AYDMTQTPASVEVAVGGTVTIKCQASESISTYLAWYQQKPGQPPKLLIYRASTLASGVSSRFKGSGSGTQFTLTISGVECADAATYYCQQGYSYSNVDNAFGGGTEVVVKR (SEQ ID NO: 862).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab23 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 870).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:844, SEQ ID NO:846 and SEQ ID NO:848, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQ ID NO: 841 or it contains the variable heavy chain sequence of SEQ ID NO: 842; and/or it further contains SEQ ID NO: 864, SEQ ID NO: 866 and SEQ ID NO: One, two or three of the polypeptide sequences of 868, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequences of SEQ ID NO: 861 or the variable light chain sequences containing SEQ ID NO: 862; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:843, SEQ ID NO:845, SEQ ID NO:847 and SEQ ID NO:849, which correspond to SEQ ID NO: the heavy chain sequence of 841 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:842; and/or one of the polypeptide sequences of SEQ ID NO:863, SEQ ID NO:865, SEQ ID NO:867 and SEQ ID NO:869 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 861 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 862; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 841 or SEQ ID NO: 842 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 861 or SEQ ID NO: 862 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:844, SEQ ID NO:846 and SEQ ID NO:848 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 841 or the variable heavy chain sequence of SEQ ID NO: 842; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:864, SEQ ID NO:866 and SEQ ID NO:868 or three, which correspond to the light chain sequence of SEQ ID NO:861 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO:862; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO:843, SEQ ID NO:845, SEQ ID NO:847 and SEQ ID NO:849 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO:841 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:842; Or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO:863, SEQ ID NO:865, SEQ ID NO:867 and SEQ ID NO:869 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:861 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:862; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO:842 variable heavy chain region; the variable light chain region of SEQ ID NO:862; the complementarity determining region (SEQ ID NO:844, SEQ ID NO:846 and SEQ ID NO:848) of the variable heavy chain region of SEQ ID NO:842; and the variable of SEQ ID NO:862 the complementarity determining region of the light chain region (SEQ ID NO:864, SEQ ID NO:866 and SEQ ID NO:868); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO:842 Variable heavy chain region; variable light chain region of SEQ ID NO:862; framework region of variable heavy chain region of SEQ ID NO:842 (SEQ ID NO:843, SEQ ID NO:845, SEQ ID NO:847 and SEQ ID NO:849); and SEQ ID NO:862 The framework regions of the variable light chain region (SEQ ID NO: 863, SEQ ID NO: 865, SEQ ID NO: 867 and SEQ ID NO: 869).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab23 comprising or consisting of SEQ ID NO: 841 and SEQ ID NO: 861; or an antibody comprising the CDRs of Ab23 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab23 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab23; or an antibody that binds to an epitope on HGF that is identical or overlaps with Ab23 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Concerning antibody Ab23, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 842 and the variable light chain sequence of SEQ ID NO: 862 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:842 and/or SEQ ID NO:862 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab23. In another embodiment of the present invention, anti-HGF antibodies such as Ab23 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab23 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab24

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：EVQLVESGGGLVQPGGSLRLSCAASGFTVGRYYMSWVRQAPGKGLEWIGIIYTHGVNPDYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVGGFNDYSDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:881)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: EVQLVESGGGLVQPGGSLRLSCAASGFTVGRYYMSWVRQAPGKGLEWIGIIYTHGVNPDYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVGGFNDYSDIWGQGTLVTVSS (SEQ ID NO: 882).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab24相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 890).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：DIQMTQSPSSLSASVGDRVTITCQASESISTYLAWYQQKPGKVPKLLIYRASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQQGYSYSNVDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:901)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: DIQMTQSPSSLSASVGDRVTITCQASESISTYLAWYQQKPGKVPKLLIYRASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQQGYSYSNVDNAFGGGTKVEIKR (SEQ ID NO: 902).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab24 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 910).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:884, SEQ ID NO:886 and SEQ ID NO:888, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQ ID NO: 881 or it contains the variable heavy chain sequence of SEQ ID NO: 882; and/or it further contains SEQ ID NO: 904, SEQ ID NO: 906 and SEQ ID NO: One, two or three of the polypeptide sequences of 908, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 901 or the variable light chain sequence containing SEQ ID NO: 902; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:883, SEQ ID NO:885, SEQ ID NO:887 and SEQ ID NO:889, which correspond to SEQ ID NO: the heavy chain sequence of 881 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:882; and/or one of the polypeptide sequences of SEQ ID NO:903, SEQ ID NO:905, SEQ ID NO:907 and SEQ ID NO:909 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 901 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 902; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 881 or SEQ ID NO: 882 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 901 or SEQ ID NO: 902 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:884, SEQ ID NO:886 and SEQ ID NO:888 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 881 or the variable heavy chain sequence of SEQ ID NO: 882; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 904, SEQ ID NO: 906 and SEQ ID NO: 908 or three, which correspond to the light chain sequence of SEQ ID NO:901 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO:902; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO:883, SEQ ID NO:885, SEQ ID NO:887 and SEQ ID NO:889 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO:881 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:882; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 903, SEQ ID NO: 905, SEQ ID NO: 907 and SEQ ID NO: 909 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:901 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:902; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO:882 variable heavy chain region; the variable light chain region of SEQ ID NO:902; the complementarity determining region (SEQ ID NO:884, SEQ ID NO:886 and SEQ ID NO:888) of the variable heavy chain region of SEQ ID NO:882; and the variable of SEQ ID NO:902 the complementarity determining region of the light chain region (SEQ ID NO: 904, SEQ ID NO: 906 and SEQ ID NO: 908); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO:882 Variable heavy chain region; variable light chain region of SEQ ID NO:902; framework region of variable heavy chain region of SEQ ID NO:882 (SEQ ID NO:883, SEQ ID NO:885, SEQ ID NO:887 and SEQ ID NO:889); and SEQ ID NO:902 The framework region of the variable light chain region (SEQ ID NO: 903, SEQ ID NO: 905, SEQ ID NO: 907 and SEQ ID NO: 909).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab24 comprising or consisting of SEQ ID NO: 881 and SEQ ID NO: 901; or an antibody comprising the CDRs of Ab24 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab24 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab24; or an antibody that binds to an epitope on HGF that is identical or overlaps with Ab24 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Ab24, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 882 and the variable light chain sequence of SEQ ID NO: 902 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:882 and/or SEQ ID NO:902 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab24. In another embodiment of the invention, anti-HGF antibodies such as Ab24 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab24 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab25

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSLEESGGRLVTPGTPLTLTCTASGFSLSSYAMGWFRQAPGKGLEWIAYIFASGSTYYASWAKGRFTISKTSTTVELKITSLTTEDTATYFCARGSGARFFPNYFAIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:921)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSLEESGGRLVTPGTPLTLTCTASGFSLSSYAMGWFRQAPGKGLEWIAYIFASGSTYYASWAKGRFTISKTSTTVELKITSLTTEDTATYFCARGSGARFFPNYFAIWGPGTLVTVSS (SEQ ID NO: 922).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab25相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 930).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：QVLTQTASSVSAAVGGTVTISCQSSQSVTNNNDLAWYQQKPGQPPKLLIYQASKLASGVPSRFKGSGSGTQFTLTISDLECDDAATYYCQGSYSGGICAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:941)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: QVLTQTASSVSAAVGGTVTISCQSSQSVTNNNDLAWYQQKPGQPPKLLIYQASKLASGVPSRFKGSGSGTQFTLTISDLECDATYYCQGSYSGGICAFGGGTEVVVKR (SEQ ID NO: 942).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab25 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 950).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:924, SEQ ID NO:926 and SEQ ID NO:928, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQIDNO:921 or it contains the variable heavy chain sequence of SEQIDNO:922; and/or it further contains SEQIDNO:944, SEQIDNO:946 and SEQIDNO: One, two or three of the polypeptide sequences of 948, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 941 or the variable light chain sequences containing SEQ ID NO: 942; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:923, SEQ ID NO:925, SEQ ID NO:927 and SEQ ID NO:929, which correspond to SEQ ID NO: the heavy chain sequence of 921 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:922; and/or one of the polypeptide sequences of SEQ ID NO:943, SEQ ID NO:945, SEQ ID NO:947 and SEQ ID NO:949 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 941 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 942; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 921 or SEQ ID NO: 922 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 941 or SEQ ID NO: 942 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 924, SEQ ID NO: 926 and SEQ ID NO: 928 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 921 or the variable heavy chain sequence of SEQ ID NO: 922; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO:944, SEQ ID NO:946 and SEQ ID NO:948 or three, which correspond to the light chain sequence of SEQ ID NO:941 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO:942; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 923, SEQ ID NO: 925, SEQ ID NO: 927 and SEQ ID NO: 929 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO: 921 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO: 922; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 943, SEQ ID NO: 945, SEQ ID NO: 947 and SEQ ID NO: 949 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:941 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:942; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: variable heavy chain region; the variable light chain region of SEQ ID NO:942; the complementarity determining region (SEQ ID NO:924, SEQ ID NO:926 and SEQ ID NO:928) of the variable heavy chain region of SEQ ID NO:922; and the variable of SEQ ID NO:942 the complementarity determining region of the light chain region (SEQ ID NO:944, SEQ ID NO:946 and SEQ ID NO:948); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: The variable heavy chain region; the variable light chain region of SEQ ID NO:942; the framework region of the variable heavy chain region of SEQ ID NO:922 (SEQ ID NO:923, SEQ ID NO:925, SEQ ID NO:927 and SEQ ID NO:929); and SEQ ID NO:942 The framework region of the variable light chain region (SEQ ID NO: 943, SEQ ID NO: 945, SEQ ID NO: 947 and SEQ ID NO: 949).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab25 comprising or consisting of SEQ ID NO: 921 and SEQ ID NO: 941; or an antibody comprising the CDRs of Ab25 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab25 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab25; or an antibody that binds to an epitope on HGF that is identical or overlaps with Ab25 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Concerning antibody Ab25, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 922 and the variable light chain sequence of SEQ ID NO: 942 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:922 and/or SEQ ID NO:942 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab25. In another embodiment of the invention, anti-HGF antibodies such as Ab25 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab25 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab26

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSVEESGGRLVTPGTPLTLTCTVSGFSLSTYTMNWVRQAPGKGLEYIGFISSSSSIDYVSWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARDFYADYIGGGYPYIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:961)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSVEESGGRLVTPGTPLTLTCTVSGFSLSTYTMNWVRQAPGKGLEYIGFISSSSSIDYVSWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARDFYADYIGGGYPYIWGPGTLVTVSS (SEQ ID NO: 962).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab26相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 970).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：ADVVMTQTPASVSEPVGGTVTIKCQASQSISSYLSWYQQKPGQPPKLLIYGASKLTSGVPSRFKGSGSGTEYTLTISDLECADAATYYCQSNYDIYSYAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:981)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: ADVVMTQTPASVSEPVGGTVTIKCQASQSISSYLSWYQQKPGQPPKLLIYGASKLTSGVPSRFKGSGSGTEYTLTISDLECADATYYCQSNYDIYSYAFGGGTEVVVKR (SEQ ID NO: 982).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab26 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 990).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO:964, SEQ ID NO:966 and SEQ ID NO:968, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQ ID NO: 961 or it contains the variable heavy chain sequence of SEQ ID NO: 962; and/or it further contains SEQ ID NO: 984, SEQ ID NO: 986 and SEQ ID NO: One, two or three of the polypeptide sequences of 988, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 981 or the variable light chain sequence containing SEQ ID NO: 982; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO:963, SEQ ID NO:965, SEQ ID NO:967 and SEQ ID NO:969, which correspond to SEQ ID NO: The heavy chain sequence of 961 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:962; and/or one of the polypeptide sequences of SEQ ID NO:983, SEQ ID NO:985, SEQ ID NO:987 and SEQ ID NO:989 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 981 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 982; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 961 or SEQ ID NO: 962 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 981 or SEQ ID NO: 982 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 964, SEQ ID NO: 966 and SEQ ID NO: 968 or three, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 961 or the variable heavy chain sequence of SEQ ID NO: 962; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 984, SEQ ID NO: 986 and SEQ ID NO: 988 or three, which correspond to the light chain sequence of SEQ ID NO: 981 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO: 982; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 963, SEQ ID NO: 965, SEQ ID NO: 967 and SEQ ID NO: 969 one, two, three or four, which correspond to the heavy chain sequence of SEQ ID NO: 961 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 962; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 983, SEQ ID NO: 985, SEQ ID NO: 987 and SEQ ID NO: 989 One, two, three or four, which correspond to the light chain sequence of SEQIDNO:981 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO:982; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: variable heavy chain region; the variable light chain region of SEQ ID NO:982; the complementarity determining region (SEQ ID NO:964, SEQ ID NO:966 and SEQ ID NO:968) of the variable heavy chain region of SEQ ID NO:962; and the variable of SEQ ID NO:982 the complementarity determining region of the light chain region (SEQ ID NO:984, SEQ ID NO:986 and SEQ ID NO:988); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: Variable heavy chain region; variable light chain region of SEQ ID NO:982; framework region of variable heavy chain region of SEQ ID NO:962 (SEQ ID NO:963, SEQ ID NO:965, SEQ ID NO:967 and SEQ ID NO:969); and SEQ ID NO:982 The framework regions of the variable light chain region (SEQ ID NO:983, SEQ ID NO:985, SEQ ID NO:987 and SEQ ID NO:989).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab26 comprising or consisting of SEQ ID NO: 961 and SEQ ID NO: 981; or an antibody comprising the CDRs of Ab26 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab26 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab26; or an antibody that binds to an epitope on HGF that is identical or overlaps with Ab26 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Ab26, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 962 and the variable light chain sequence of SEQ ID NO: 982 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO:962 and/or SEQ ID NO:982 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab26. In another embodiment of the present invention, anti-HGF antibodies such as Ab26 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab26 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab27

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：QSLEESGGDLVKPGGTLTLTCTASGFSFSDDHYMCWVRQAPGKGLQWIACMYVGSSGATYYASWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCARDDWTSYYAWGYWALWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:1001)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: QSLEESGGDLVKPGGTLLTTCTASGFSFSDDHYMCWVRQAPGKGLQWIACMYVGSSGATYYASWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCARDDWTSYYAWGYWALWGPGTLVTVSS (SEQ ID NO: 1002).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab27相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 1010).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：ADIVMTQNPASVSEPVGGTVTIKCQASQSVNSWLSWYQQKPGQPPKFLIYKASTLASGVSSRFKGSGIGTEFTLTISDLECADAATYYCQFSNSGTIYGSGFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:1021)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: ADIVMTQNPASVSEPVGGTVTIKCQASQSVNSWLSWYQQKPGQPPKFLIYKASTLASGVSSRFKGSGIGTEFTLTISDLECADAATYYCQFSNSGTIYGSGFGGGTEVVVKR (SEQ ID NO: 1022).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab27 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 1030).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO: 1004, SEQ ID NO: 1006 and SEQ ID NO: 1008, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQ ID NO: 1001 or it contains the variable heavy chain sequence of SEQ ID NO: 1002; and/or it further contains SEQ ID NO: 1024, SEQ ID NO: 1026 and SEQ ID NO: One, two or three of the polypeptide sequences of 1028, which correspond to the complementarity determining region (CDR or hypervariable region) of the light chain sequence of SEQ ID NO: 1021 or the variable light chain sequence containing SEQ ID NO: 1022; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO: 1003, SEQ ID NO: 1005, SEQ ID NO: 1007 and SEQ ID NO: 1009, which correspond to SEQ ID NO: The heavy chain sequence of 1001 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO:1002; and/or one of the polypeptide sequences of SEQ ID NO:1023, SEQ ID NO:1025, SEQ ID NO:1027 and SEQ ID NO:1029 , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 1021 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 1022; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 1001 or SEQ ID NO: 1002 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 1021 or SEQ ID NO: 1022 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 1004, SEQ ID NO: 1006 and SEQ ID NO: 1008 or three, which correspond to the heavy chain sequence of SEQ ID NO: 1001 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 1002; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 1024, SEQ ID NO: 1026 and SEQ ID NO: 1028 or three, which correspond to the light chain sequence of SEQ ID NO: 1021 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO: 1022; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the present invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of the following: one of the polypeptide sequences of SEQ ID NO: 1003, SEQ ID NO: 1005, SEQ ID NO: 1007 and SEQ ID NO: 1009 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO: 1001 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO: 1002; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 1023, SEQ ID NO: 1025, SEQ ID NO: 1027 and SEQ ID NO: 1029 One, two, three or four, which correspond to the light chain sequence of SEQIDNO: 1021 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO: 1022; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the present invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO: 1002 variable heavy chain region; the variable light chain region of SEQ ID NO: 1022; the complementarity determining region (SEQ ID NO: 1004, SEQ ID NO: 1006 and SEQ ID NO: 1008) of the variable heavy chain region of SEQ ID NO: 1002; and the variable of SEQ ID NO: 1022 the complementarity determining region of the light chain region (SEQ ID NO: 1024, SEQ ID NO: 1026 and SEQ ID NO: 1028); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the present invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO: 1002 variable heavy chain region; variable light chain region of SEQ ID NO: 1022; framework regions (SEQ ID NO: 1003, SEQ ID NO: 1005, SEQ ID NO: 1007 and SEQ ID NO: 1009) of the variable heavy chain region of SEQ ID NO: 1002; and SEQ ID NO: 1022 The framework region of the variable light chain region (SEQ ID NO: 1023, SEQ ID NO: 1025, SEQ ID NO: 1027 and SEQ ID NO: 1029).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab27 comprising or consisting of SEQ ID NO: 1001 and SEQ ID NO: 1021; or an antibody comprising the CDRs of Ab27 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab27 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab27; or an antibody that binds to an epitope on HGF that is identical or overlaps with Ab27 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Ab27, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 1002 and the variable light chain sequence of SEQ ID NO: 1022 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO: 1002 and/or SEQ ID NO: 1022 which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab27. In another embodiment of the present invention, anti-HGF antibodies such as Ab27 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab27 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

Antibody Ab28

在一个实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所具有的重链序列包含以下所阐述的序列：EVQLVESGGGLVQPGGSLRLSCAASGFTVSSNAISWVRQAPGKGLEWVGVIYVIGVTDYASSAQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVYDSGWNHFNLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDARVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:1041)。

In one embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable heavy chain sequence comprising the sequence set forth below: EVQLVESGGGLVQPGGSLRLSCAASGFTVSSNAISWVRQAPGKGLEWVGVIYVIGVTDYASSAQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVYDSGWNHFNLWGQGTLVTVSS (SEQ ID NO: 1042)

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其具有与Ab28相同的表位特异性且其所含恒定重链序列包含以下所阐述的序列：ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDARVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK( SEQ ID NO: 1050).

在另一实施方案中，本发明包括具有针对HGF的结合特异性的抗体和抗体片段，其所含轻链序列包含以下所阐述的序列：DIQMTQSPSTLSASVGDRVTITCQASQSISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQAYSVANVDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:1061)。

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF comprising a variable light chain sequence comprising the sequence set forth below: DIQMTQSPSTLSASVGDRVTITCQASQSISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQAYSVANVDNAFGGGTKVEIKR (SEQ ID NO: 1062).

In another embodiment, the invention includes antibodies and antibody fragments having binding specificity for HGF that bind the same epitope as Ab28 and that contain a constant light chain sequence comprising the sequence set forth below: TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 1070).

In another embodiment, the present invention includes antibodies and antibody fragments having binding specificity for HGF comprising one, two or three of the polypeptide sequences of SEQ ID NO: 1044, SEQ ID NO: 1046 and SEQ ID NO: 1048, It corresponds to the complementarity determining region (CDR or hypervariable region) of the heavy chain sequence of SEQ ID NO: 1041 or it contains the variable heavy chain sequence of SEQ ID NO: 1042; and/or it further contains SEQ ID NO: 1064, SEQ ID NO: 1066 and SEQ ID NO: One, two or three of the polypeptide sequences of 1068, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 1061 or the variable light chain sequences containing SEQ ID NO: 1062; or An antibody or fragment comprising a sequence combination that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to said polypeptide sequence. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of the exemplified variable heavy chain and variable light chain sequences or the heavy chain and light chain sequences set forth above or in combination with A combination of one or more of the sequences that are at least 90% or 95% identical.

The invention also encompasses anti-HGF antibodies and antibody fragments comprising one, two, three or four of the polypeptide sequences of SEQ ID NO: 1043, SEQ ID NO: 1045, SEQ ID NO: 1047 and SEQ ID NO: 1049, which correspond to SEQ ID NO: The heavy chain sequence of 1041 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 1042; , two, three or four, which correspond to the light chain sequence of SEQ ID NO: 1061 or the framework region (FR or constant region) of the variable light chain sequence of SEQ ID NO: 1062; or these polypeptide sequences or at least 80% thereof, Combinations of sequences that are 90% or 95% identical.

In another embodiment of the invention, the antibodies and antibody fragments or fragments thereof of the invention comprise or consist of the FR, CDR, variable heavy and variable light chain sequences and the heavy and light chain sequences set forth above. A combination of one or more of the strand sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment of the present invention, the anti-HGF antibody fragment of the present invention comprises or consists of the polypeptide sequence of SEQ ID NO: 1041 or SEQ ID NO: 1042 or a polypeptide at least 90% or 95% identical thereto. In another embodiment of the invention, the antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 1061 or SEQ ID NO: 1062 or a polypeptide at least 90% or 95% identical thereto.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 1044, SEQ ID NO: 1046 and SEQ ID NO: 1048 or three, which correspond to the heavy chain sequence of SEQ ID NO: 1041 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 1042; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one or both of the polypeptide sequences of SEQ ID NO: 1064, SEQ ID NO: 1066 and SEQ ID NO: 1068 or three, which correspond to the light chain sequence of SEQ ID NO: 1061 or the complementarity determining region (CDR or hypervariable region) of the variable light chain sequence of SEQ ID NO: 1062; or at least 90% or 95% identical to said polypeptide sequence sequence.

In another embodiment of the invention, the antibody or antibody fragment having binding specificity for HGF comprises or consists of one of the polypeptide sequences of SEQ ID NO: 1043, SEQ ID NO: 1045, SEQ ID NO: 1047 and SEQ ID NO: 1049 Either, both, three or four, it corresponds to the heavy chain sequence of SEQIDNO: 1041 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO: 1042; or at least 90% with the polypeptide sequence or 95% identical sequences.

In another embodiment of the invention, the subject antibody or antibody fragment having binding specificity for HGF comprises or consists of: in the polypeptide sequences of SEQ ID NO: 1063, SEQ ID NO: 1065, SEQ ID NO: 1067 and SEQ ID NO: 1069 One, two, three or four, which correspond to the light chain sequence of SEQIDNO: 1061 or the framework region (FR or constant region) of the variable light chain sequence of SEQIDNO: 1062; or at least 90 to the polypeptide sequence % or 95% identical sequences.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO: 1042 The variable heavy chain region; the variable light chain region of SEQ ID NO: 1062; the complementarity determining region (SEQ ID NO: 1044, SEQ ID NO: 1046 and SEQ ID NO: 1048) of the variable heavy chain region of SEQ ID NO: 1042; and the variable of SEQ ID NO: 1062 the complementarity determining region of the light chain region (SEQ ID NO: 1064, SEQ ID NO: 1066 and SEQ ID NO: 1068); or a sequence at least 90% or 95% identical thereto.

The invention also encompasses antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the invention, the antibody fragment having binding specificity for HGF comprises or consists of one, two, three or more (including all) of the following antibody fragments: SEQ ID NO: 1042 variable heavy chain region; variable light chain region of SEQ ID NO: 1062; framework region (SEQ ID NO: 1043, SEQ ID NO: 1045, SEQ ID NO: 1047 and SEQ ID NO: 1049) of the variable heavy chain region of SEQ ID NO: 1042; and SEQ ID NO: 1062 The framework region of the variable light chain region (SEQ ID NO: 1063, SEQ ID NO: 1065, SEQ ID NO: 1067 and SEQ ID NO: 1069).

In a particularly preferred embodiment of the present invention, the anti-HGF antibody is Ab28 comprising or consisting of SEQ ID NO: 1041 and SEQ ID NO: 1061; or an antibody comprising the CDRs of Ab28 and having at least one of the biological activities set forth herein or an antibody fragment; or an anti-HGF antibody that competes with Ab28 for binding to HGF, preferably an antibody that contains a sequence that is at least 90% or 95% identical to the sequence of Ab28; or an antibody that binds to an epitope on HGF that is identical or overlapping with Ab28 .

In another particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Ab28, the Fab fragment preferably comprises the variable heavy chain sequence of SEQ ID NO: 1042 and the variable light chain sequence of SEQ ID NO: 1062 or a sequence at least 90% or 95% identical thereto. This embodiment of the invention also includes Fabs comprising additions, deletions and variants of SEQ ID NO: 1042 and/or SEQ ID NO: 1062, which retain binding specificity for HGF.

In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab28. In another embodiment of the present invention, anti-HGF antibodies such as Ab28 or Fab fragments thereof can be obtained via expression in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems such as yeast cells (e.g. haploid or diploid yeast, such as haploid or diploid Pichia) and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

In another embodiment, the present invention also relates to a polynucleotide encoding an antibody polypeptide with binding specificity for HGF, which includes the heavy chain and/or light chain of Ab28 and the FR, CDR, variable heavy chain as set forth above. Fragments, variants, combinations of one or more of chain and variable light chain sequences and heavy chain and light chain sequences, including all said sequences or sequences at least 90% or 95% identical thereto.

In another embodiment, the invention encompasses an isolated anti-HGF antibody comprising any of the previously identified _VH polypeptide sequences; and further comprising any of the previously identified _VL polypeptide sequences or, wherein one or more of the framework residues (FR residues) in the _VH or _VL polypeptide has been substituted with another amino acid residue to generate an anti-HGF antibody that specifically binds HGF. Humanized and chimeric forms of these antibodies are encompassed by the invention. Chimeric antibodies may comprise antibodies derived from IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, IgG7, IgG8, IgG9, IgG10, IgG11, IgG12, IgG13, IgG14, IgG15, IgG16, IgG17, IgG18 or IgG19, IgG20, IgG21 and others The Fc of the antibody constant region, which optionally may be mutagenized or fragmented.

In one embodiment of the invention, said antibody or _VH or _VL polypeptide is derived from or selected from one or more rabbit B cell populations, followed by the humanization process mentioned herein.

In another embodiment of the invention, anti-HGF antibodies and fragments thereof having binding specificity for human HGF and allelic variants of human HGF can also bind to non-human HGF polypeptides, including primate species of human HGF Animal homologues, such as HGF expressed in chimpanzees, monkeys, apes, gigantic apes, gorillas, lemurs, and other primates; and possibly bind to non-primate HGF polypeptides, such as rabbit, canine Animal, rodent, feline and other HGF species homologues.

In another embodiment of the invention, the anti-HGF antibodies and fragments thereof do not have binding specificity for HGFR (c-met). In another embodiment of the invention, anti-HGF antibodies and fragments thereof inhibit the binding of HGF to HGFR. In another embodiment of the present invention, anti-HGF antibodies and fragments thereof inhibit the binding of HGF to HGFR and/or its multimers and/or antagonize their biological effects.

In addition, the anti-HGF antibodies and fragments thereof of the invention may be post-translationally modified to add effector moieties, such as chemical linkers; detectable moieties, such as fluorescent dyes, enzymes, substrates, bioluminescent substances, radioactive substances, and chemiluminescent moieties; or functional Moieties such as streptavidin, avidin, biotin, cytotoxins, cytotoxic agents and radioactive substances.

With regard to detectable moieties, other exemplary enzymes include, but are not limited to, horseradish peroxidase, acetylcholinesterase, alkaline phosphatase, beta-galactosidase, and luciferase. Other exemplary fluorescent substances include, but are not limited to, rhodamine, fluorescein, fluorescent isothiocyanate, umbelliferone, dichlorotriazinylamine, phycoerythrin, and dansyl chloride. Other exemplary chemiluminescent moieties include, but are not limited to, luminol. Other exemplary bioluminescent substances include, but are not limited to, luciferin and luminescent proteins. Other exemplary radioactive materials include, but are not limited to, iodine 125 ( ¹²⁵ I), carbon 14 ( ¹⁴ C), sulfur 35 ( ³⁵ S), tritium ( ³ H), and phosphorus 32 ( ³² P).

In another embodiment, anti-HGF antibodies and fragments of the invention may be linked to one or more other functional moieties or administered in combination therapy, exemplary cytotoxic agents include, but are not limited to, methotrexate, aminopterin, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine; alkylating agents such as methylenedichlorodiethylamine, thioepachlorambucil, Melphalan, Carmustine (BSNU), Mitomycin C, Lomustine (CCNU), 1-Methylnitrosourea, Cyclothionamide, Methyldichlorodiethylamine, Busulfan , dibromomannitol, streptozotocin (streptozotocin), mitomycin C, cis-dichlorodiamine platinum (II) (DDP) cisplatin and carboplatin (Birdine); anthracyclines ( anthracyclines), including daunorubicin (formerly daunomycin), doxorubicin (adriamycin), detorubicin, carminomycin, idarubicin, Epirubicin, mitoxantrone, and bisantrene; antibiotics, including dactinomycin (actinomycin D), bleomycin, calicheamicin, mithramycin and anthramycin (AMC); and antimitotic agents such as vinca alkaloids, vincristine and vinblastine. Other cytotoxic agents include paclitaxel (taxol), ricin, pseudomonas exotoxin, gemcitabine, cytochalasin B, gramicidin D, ethidium bromide, emetine, etoposide, Tenoposide, colchicine, dihydroxyanthraxin diketone, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine ( lidocaine), propranolol, puromycin, procarbazine, hydroxyurea, asparaginase, corticosteroids, mytotane (O,P'-(DDD)), Interferons and mixtures of these cytotoxic agents.

Other cytotoxic agents include, but are not limited to, chemotherapeutic agents such as carboplatin, cisplatin, paclitaxel, gemcitabine, calicheamicin, doxorubicin, 5-fluorouracil, mitomycin C, actinomycin D , cyclophosphamide, vincristine and bleomycin. Toxic proteins from plants and bacteria, such as ricin, diphtheria, and pseudomonad toxins, can be conjugated to humanized or chimeric antibodies or binding fragments thereof to generate cell-type specific lethal agents (Youle et al. USA77:5483 (1980); Gilliland et al., Proc.Nat'1Acad.Sci.USA77:4539 (1980); Krolick et al., Proc.Nat'1Acad.Sci.USA77:5419 (1980)).

Other cytotoxic agents that may be used in combination with the anti-HGF antibodies and fragments of the invention include cytotoxic ribonucleases as described by Goldenberg in US Patent No. 6,653,104. Embodiments of the invention also pertain to radioimmunoconjugates wherein an alpha or beta particle emitting radionuclide is stably coupled to an antibody or binding fragment thereof with or without the use of a complex forming agent. Such radionuclides include beta emitters such as phosphorus-32 ( ³² P), scandium-47 ( ⁴⁷ Sc), copper-67 ( ⁶⁷ Cu), gallium-67 ( ⁶⁷ Ga), yttrium-88 ( ⁸⁸ Y) , yttrium-90 ( ⁹⁰ Y), iodine-125 ( ¹²⁵ I), iodine-131 ( ¹³¹ I), samarium-153 ( ¹⁵³ Sm), lutetium-177 ( ¹⁷⁷ Lu), rhenium-186 ( ¹⁸⁶ Re) or rhenium -188 ( ¹⁸⁸ Re); and alpha emitters such as astatine-211 ( ²¹¹ At), lead-212 ( ²¹² Pb), bismuth-212 ( ²¹² Bi) or bismuth-213 ( ²¹³ Bi) or actinium-225 ( ²²⁵ Ac). The term "cytotoxic agent" as used herein broadly includes any substance that inhibits or prevents cellular function and/or causes cellular destruction and "chemotherapeutic agents" useful in the treatment of cancer. The term is intended to include radioactive isotopes; chemotherapeutic agents such as methotrexate, doxorubicin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan , mitomycin C, chlorambucil, daunomycin or other intercalating agents; enzymes and fragments thereof such as nucleolytic enzymes, antibiotics; and toxins such as small molecule toxins of bacterial, fungal, plant or animal origin or enzyme active toxins, including fragments and/or variants thereof, and various antitumor or anticancer agents disclosed below. Other cytotoxic agents are described below. Tumorcidal agents cause the destruction of tumor cells.

A "chemotherapeutic agent" is a compound useful in the treatment of cancer. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan, and pipoxulfan; aziridines such as benzotepa, carboquinone, methutepa, and uretipa; Ethyleneimines and methylmelamines, including hexamethylmelamine, triptamide, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylmelamine; ); camptothecin (including the synthetic analogs topotecan); bryostatin; calistatin; CC-1065 (including its synthetic analogs adolaisine, kazelesine, and bizelesine); Rosary Cyclopeptides (specifically, Nostoc cyclic peptide 1 and Nostoc cyclic peptide 8); Aplysia spp.; Duocarcinol (including the synthetic analogues KW-2189 and CB1-TM1); Cortinol; Alkalis; crotonol; spongostatin; nitrogen mustards such as chlorambucil, naphthalene, chlorphosphamide, estramustine, ifosfamide, methylenedichlorodiethylamine, methylenedichlorodiethylamine Ethylamine oxide hydrochloride, melphalan, nemethambucil, cholesteryl phenylacetate mustard, prednimustine, trofosamide, uracil mustard; nitrosoureas such as carmustine, chloride Ureacin, fomustine, lomustine, nimustine, and ramustine; antibiotics such as enediyne antibiotics (eg, calicheamicins, especially calicheamicin gamma and calicheamicin omega (See, eg, Agnew, ChemIntl. Ed. Engl., 33:183-186 (1994)); Danemycins, including Danemycin A; Bisphosphonates, such as clodronate; Espera and neocarcinogen chromophores and related chromoprotein enediyne antibiotic chromophores), aclarithromycin, actinomycin, anthraninomycin, azaserine, bleomycin, actinomycin C, Carrubicin, Carrubicin, Carcinophilin, Chromomycin, Dactinomycin, Daunomycin, Detorubicin, 6-diazo-5-oxo-L-Zhengliang amino acid, Doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolinyl-doxorubicin, and deoxydoxorubicin), epirubicin, Sobicin, idarubicin, moxicilomycin, mitomycin (such as mitomycin C), mycophenolic acid, nogamycin, olivine, pelomycin, phenenomycin , puromycin, quinamycin, rhodorubicin, streptomycin, streptozocin, tuberculin, ubenimex, netastatin, zorubicin; antimetabolites such as ammonia Methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as dinotrazine, methotrexate, pteroxin, trimezat; purine analogs such as fludarabine, 6-mercaptopurine , thiametidine, thioguanine; pyrimidine analogs such as cyclocitidine, azacitidine, 6-azuridine, carmofur, cytarabine, dideoxyuridine, deoxyfluridine, enoxa Tabine, floxuridine; androgens, such as calasterone, drostanolone propionate, cyclic thiosterol, metrostan, testolactone; antiadrenergics, such as aminoglutethimide, mitotane, trirolactone Stein; folic acid supplements such as folinic acid; acetglucuronolactone; aldophosphamide glucoside; aminolevulinic acid; eniluracil; amsacrine; besbucil; bisantrene; edatrexate; Amides; colcemid; decacrine; ilonide; etricetium; epothilone; ethoxydine; gallium nitrate; hydroxyurea; mushroom polysaccharides; Densine and ansamectin; Mitoguanidine hydrazone; Mitoxantrone; Mopedadol; Diamine nitroacridine; Pentostatin; Methionine; Pirarubicin; Loxoantrone; Podophyllic acid ; 2-acetylhydrazide; Procarbazine; Polysaccharide complex (JHS Natural Products, Eugene, Oreg.); Razoxane; Rhizopus; Cizoran; Trichothecenes (especially T-2 toxin, mucin A, bacitracin A, and serpentine); urethane; vindesine; dacarbazine; mannomustine; dibromomannitol; Bromodulcitol; Pipobromide; Gacitocin; Cytarabine ("Ara-C");Cyclophosphamide;Thiotepa; Taxoids, eg Paclitaxel (Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE.TM. Polyoxyethylated castor oil-free albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.) and docetaxel (Rhone-Poulenc Rorer, Antony, France); chlorambucil; Gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone ; vincristine; Vinorelbine; Nosolin; Teniposide; Edatrexate; Daunorubicin; Amopterin; Xeloda; Ibandronate; CPT-11; Topoisomerase inhibitor RFS2000; II Fluoromethylornithine (DMFO); retinoids, such as retinoic acid; capecitabine; and pharmaceutically acceptable salts, acids, or derivatives of any of the above. Also included in the definition of "chemotherapeutic agent" above are antihormonal agents that act to modulate or inhibit the effects of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen ( include Tamoxifen), Raloxifene, Triproxifen, 4-Hydroxytamoxifen, Travoxifen, Naloxifen, LY117018, Onapristone, and FARESTON. Toremifene; Aroma Suppressant Aromatase inhibitors of enzymes that regulate estrogen production in the adrenal gland, such as 4(5)-imidazole, aminoglutethimide, megestrol acetate, Exemestane, Formestane, Fadrozole, Vorozole, Letrozole and Anastrozole; and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and troxatabine (a 1,3-dioxane pentane nucleoside cytosine analogs); antisense oligonucleotides, specifically those that inhibit the expression of genes in signaling pathways involved in abnormal cell proliferation, such as PKC-α, Ralf and H - Ras; ribonucleases, such as VEGF expression inhibitors (e.g. ribonuclease) and HER2 expression inhibitors; vaccines, such as gene therapy vaccines, e.g. vaccine, vaccines and vaccine; rIL-2; Topoisomerase 1 inhibitors; rmRH; and a pharmaceutically acceptable salt, acid or derivative of any of the above. Preferred examples of cytotoxic agents used in combination with the anti-HGF antibodies and fragments of the invention are erlotinib, gemcitabine, pemetrexole, docetaxel, Folfox chemotherapy regimen combination, paclitaxel and bevacizumab.

Other cytotoxic agents include cytotoxic ribonucleases as described by Goldenberg in US Patent No. 6,653,104. Embodiments of the invention also pertain to radioimmunoconjugates wherein an alpha or beta particle emitting radionuclide is stably coupled to an antibody or binding fragment thereof with or without the use of a complex forming agent. Such radionuclides include beta emitters such as phosphorus-32 ( ³² P), scandium-47 ( ^47S c), copper-67 ( ⁶⁷ Cu), gallium-67 ( ⁶⁷ Ga), yttrium-88 ( ⁸⁸ Y) , yttrium-90 ( ⁹⁰ Y), iodine-125 ( ¹²⁵ I), iodine-131 ( ¹³¹ I), samarium-153 ( ¹⁵³ Sm), lutetium-177 ( ¹⁷⁷ Lu), rhenium-186 ( ¹⁸⁶ Re) or rhenium -188 ( ¹⁸⁸ Re); and alpha emitters such as astatine-211 ( ²¹¹ At), lead-212 ( ²¹² Pb), bismuth-212 ( ²¹² Bi) or bismuth-213 ( ²¹³ Bi) or actinium-225 ( ²²⁵ Ac).

Methods of conjugating antibodies or binding fragments thereof to detectable moieties and analogs thereof are known in the art, such as by Hunter et al., Nature 144:945 (1962); David et al., Biochemistry 13:1014 (1974); Pain et al., J. Immunol. Meth. 40:219 (1981); and those methods described by Nygren, J., Histochem. and Cytochem. 30:407 (1982).

Embodiments described herein also include variants and equivalents substantially homologous to the antibodies, antibody fragments, diabodies, SMIPs, camelid antibodies, Nanobodies, IgNARs, polypeptides, variable regions and CDRs described herein. things. These variants and equivalents may contain, for example, conservative substitution mutations (ie, one or more amino acids are substituted with similar amino acids). For example, a conservative substitution refers to the substitution of one amino acid with another within the same general class, such as one acidic amino acid with another acidic amino acid, one basic amino acid with another basic amino acid, or a A neutral amino acid is substituted with another neutral amino acid. The intent of conservative amino acid substitutions is well known in the art.

In another embodiment, the invention encompasses polypeptide sequences having at least 90% or more sequence homology to any one or more of the polypeptide sequences of antibody fragments, variable regions and CDRs set forth herein. More preferably, the invention encompasses at least 95% or more sequence homology to any one or more of the polypeptide sequences of antibody fragments, variable regions and CDRs set forth herein, even more preferably at least 98% or A polypeptide sequence with the above sequence homology, and still more preferably at least 99% or above sequence homology. Methods for determining homology between nucleic acid and amino acid sequences are well known to those of ordinary skill.

In another embodiment, the present invention also covers the above-mentioned polypeptide homologues of the antibody fragments, variable regions and CDRs described herein that further have anti-HGF activity. Non-limiting examples of anti-HGF activity are set forth herein.

In another embodiment, the present invention also encompasses the production and use of anti-idiotypic antibodies that bind any of the above sequences. In an exemplary embodiment, such anti-idiotypic antibodies can be administered to a subject who has received anti-HGF antibodies, thereby modulating, reducing or neutralizing the effect of the anti-HGF antibodies. Such anti-idiotypic antibodies may also be useful in the treatment of autoimmune diseases that are characterized by the presence of anti-HGF antibodies. Another exemplary use of such anti-idiotypic antibodies is for detection of anti-HGF antibodies of the invention, eg, monitoring the amount of anti-HGF antibodies present in blood or other body fluids of a subject.

The invention also encompasses anti-HGF antibodies comprising any of the polypeptide or polynucleotide sequences described herein in place of any of the other polynucleotide sequences described herein. For example, the invention encompasses antibodies comprising a combination of any of the variable light chain and variable heavy chain sequences described herein, and also encompasses substitutions of any of the CDR sequences described herein Antibodies generated by any of the other CDR sequences described above, but are not limited thereto.

Additional Exemplary Embodiments of the Invention

In another embodiment, the invention encompasses one or more anti-human HGF antibodies or antibody fragments that specifically bind to the same linear or conformational epitope on an intact human HGF polypeptide or a fragment thereof as an anti-human HGF antibody and/or or compete for binding to the same linear or conformational epitope, the anti-human HGF antibody is selected from the group consisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13Ab14, Ab15, Ab16 , Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28. In a preferred embodiment, the anti-human HGF antibody or fragment specifically binds to a complete human HGF polypeptide or a fragment thereof with Ab1, Ab2, Ab7, Ab8, Ab9, Ab10, Ab12, Ab14, Ab19, Ab21, Ab23, Ab24, Ab25 and Ab28 share the same linear or conformational epitope and/or compete for binding to the same linear or conformational epitope.

In another embodiment of the present invention, the specific binding to the complete HGF polypeptide or its fragments consists of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Anti-human HGF antibodies to the same linear or conformational epitope to which Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27, and Ab28 specifically bind bind to by epitope mapping, using spanning native human HGF The HGF epitope is determined by overlapping linear peptide fragments of the full length of the polypeptide.

The present invention also relates to anti-HGF antibodies that bind to the same HGF epitope as the antibodies or antibody fragments disclosed herein and/or compete with anti-HGF antibodies for binding to HGF, including (but not limited to) selected from the group consisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13Abl4, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28 or preferably Ab1, An anti-HGF antibody of one of Ab2, Ab7, Ab8, Ab9, Ab10, Ab12, Ab14, Ab19, Ab21 , Ab23, Ab24, Ab25 or Ab28. In another embodiment, the present invention also relates to an isolated anti-HGF antibody or antibody fragment comprising an antibody selected from the group consisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13Ab14 , Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28 or preferably such as Ab1, Ab2, Ab7, Ab8, Ab9, Ab10, Ab12, Ab14, Ab19, Ab21, One or more of the CDRs contained in the VH polypeptide of the anti-HGF antibody of one of Ab23, Ab24, Ab25 and Ab28.

In one embodiment of the invention, the anti-human HGF antibody comprises at least 2 complementarity determining regions (CDRs) in each of the variable light chain and variable heavy chain regions, which are combined with an anti-human HGF antibody selected from the group consisting of anti-HGF antibodies. Those CDRs contained in an HGF antibody selected from the group consisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16 are identical , Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28 or preferably Ab1, Ab2, Ab7, Ab8, Ab9, Ab10, Ab12, Ab14, Ab19, Ab21, Ab23, Ab24, Ab25 and one of Ab28.

In a preferred embodiment, the above-discussed anti-human HGF antibody comprises at least 2 complementarity determining regions (CDRs) in each of the variable light chain and variable heavy chain regions, which are selected from the group consisting of Ab1, Ab2, Ab7, Those complementarity determining regions contained in the anti-HGF antibodies of Ab8, Ab9, Ab10, Ab12, Ab14, Ab19, Ab21, Ab23, Ab24, Ab25 and Ab28 were identical. In another embodiment, all of the CDRs of the anti-human HGF antibody discussed above are identical to the CDRs contained in an anti-human HGF antibody selected from the group consisting of an anti-HGF antibody selected from Ab1, and Ab28 or preferably as one of Ab1 , Ab2, Ab7, Ab8, Ab9, Ab10, Ab12, Ab14, Ab19, Ab21 , Ab23, Ab24, Ab25 and Ab28. In a preferred embodiment of the present invention, all CDRs of the anti-human HGF antibodies discussed above are combined with Ab1, Ab2, Ab7, Ab8, Ab9, Ab10, Ab12, Ab14, Ab19, Ab21, Ab23, Ab24, Ab25 or The CDRs contained in Ab28 or preferably in one of Ab1 , Ab2, Ab7, Ab8, Ab9, Ab10, Ab12, Ab14, Ab19, Ab21 , Ab23, Ab24, Ab25 and Ab28 are identical.

The present invention also encompasses glycosylation of one or more of the anti-human HGF antibodies discussed above; containing an Fc region that has been modified to alter effector function, half-life, proteolysis and/or glycosylation; human, human in origin single-chain or chimeric; and is a humanized antibody derived from a rabbit (parental) anti-human HGF antibody.

The invention also encompasses one or more anti-human HGF antibodies, wherein the variable light chain region and the framework regions (FR) in the variable heavy chain region of the antibody are respectively unmodified or modified with the corresponding Human FR modified by replacing FR residues with up to 2 or 3 human FR residues in the variable light or heavy chain region, and wherein the human FRs are derived from human variable heavy and light chain antibody sequences selected from the group consisting of A library of human germline antibody sequences based on a high degree of homology to the corresponding rabbit variable heavy or light chain region relative to other human germline antibody sequences contained in the human germline antibody sequence library.

In one embodiment of the invention, the anti-human HGF antibody or fragment specifically binds to HGF-expressing human cells and/or circulating soluble HGF molecules in vivo, including on human cells of patients suffering from diseases associated with HGF-expressing cells HGF expressed by or by said human cells.

In another embodiment, the disease is selected from cancer, including ovarian cancer, breast cancer, lung cancer (small cell or non-small cell), colon cancer, prostate cancer, pancreatic cancer, kidney cancer, gastric cancer, liver cancer, head and neck cancer Tumors, melanoma, sarcoma, and brain tumors in children or adults (such as glioblastoma); leukemia; lymphoma; macular degeneration; Alzheimer's disease; and malaria infection. In a preferred embodiment, the disease is selected from cancer or macular degeneration. In a particularly preferred embodiment, the disease is cancer, such as ovarian cancer, breast cancer, lung cancer (small cell or non-small cell), colon cancer, prostate cancer, pancreatic cancer, kidney cancer, gastric cancer, liver cancer, head and neck cancer Tumors, melanoma, sarcoma, and brain tumors in children or adults (such as glioblastoma); leukemia; lymphoma or another cancer.

The invention also encompasses anti-human HGF antibodies or fragments linked directly or indirectly to a detectable label or therapeutic agent.

The invention also encompasses one or more nucleic acid sequences that facilitate expression of an anti-human HGF antibody or antibody fragment as set forth above, including those comprising or consisting of yeast or human preferred codons. The invention also encompasses vectors (including plasmid or recombinant viral vectors) comprising the nucleic acid sequences. The invention also encompasses host cells or recombinant host cells expressing at least one of the antibodies set forth above, including mammalian, yeast, bacterial and insect cells. In a preferred embodiment, the host cell is a yeast cell. In another preferred embodiment, the yeast cells are diploid yeast cells. In a more preferred embodiment, the yeast cell is Pichia pastoris.

The present invention also encompasses a method of treatment comprising administering to a patient suffering from a disease or condition associated with HGF expressing cells a therapeutically effective amount of at least one anti-human HGF antibody or fragment described herein. It is also contemplated that the method of treatment may involve the administration of two or more anti-HGF antibodies or fragments thereof and are disclosed herein. If more than one antibody is administered to a patient, the multiple antibodies can be administered simultaneously or simultaneously, or can be staggered. Treatable diseases are presented in the non-limiting list set forth above and elsewhere herein. In a preferred embodiment, the disease is selected from cancer or macular degeneration. In a particularly preferred embodiment, the disease is cancer. In another embodiment, the treatment further comprises administering another therapeutic agent or regimen selected from chemotherapy, radiation therapy, cytokine administration, or gene therapy.

In a non-limiting embodiment of the invention, another therapeutic agent or regimen includes Taxol (paclitaxel) or a derivative thereof; a platinum compound such as carboplatin or cisplatin; an anthracycline such as doxorubicin; Alkylating agents such as cyclophosphamide; antimetabolites such as 5-fluorouracil or etoposide. In addition, other therapeutic agents may be used in combination with the anti-HGF antibodies and fragments of the invention, or other substances or methods for the treatment of cancer or other conditions in which it would be therapeutically advantageous to use an HGF antagonist.

For example, other substances can be radiation, chemotherapy, another anti-angiogenic agent, anti-proliferative agent. Other substances that may be used in combination with the anti-HGF antibodies and fragments of the invention are disclosed below. Preferred examples include EGFR inhibitors, such as nucleoside analogs; or platelet-derived growth factor receptor (PDGFR), vascular endothelial growth factor receptor (VEGFR), c-KIT kinase or FMS-like tyrosine kinase 3 (FLT3) Inhibitors. Such EGFR antagonists can work by binding to EGFR and competitively blocking EGF binding or by EGF activation (such as the anti-EGFR mAbs cetuximab and panitumumab) or by inhibiting the tyrosine of EGFR Acid kinase activity (such as erlotinib and gefitinib) to work. More generally, downstream signaling pathways that can be inhibited by second agents of the invention include the RAS-RAF-MEK-APK pathway and the P13K-AKT pathway. Many other signaling pathways and their inhibitors are well known to those skilled in the art of cell biology. Nucleoside analogs include gemcitabine, methotrexate, 5-fluorouracil, cytosine cytarabine, behenoylcytosine arabinoside, tegafur, UFT, and the like. Inhibitors of PDGFR, VEGFR, c-KIT kinase or FLT3 include sunitinib, sorafenib, motesanib and their analogs.

In addition, anti-HGF antibodies and fragments of the invention may be used in combination with hedgehog inhibitors, such as inhibitors of the hedgehog (HH) signaling pathway, particularly the HH pathway in humans, e.g., to inhibit the HH protein from stimulating cells via this pathway The ability of agents, also known as HH pathway inhibitors or HH inhibitors for short. Such agents may bind to HH ligands, one or more of Sonic Hedgehog (SHH), Indian Hedgehog (IHH) and Desert Hedgehog (DHH); or to Patched1 (PTCH1 ) or Patched2 ( PTCH2) receptor; or bind to downstream mediators, such as Smoothened (SMOH) or SuFu or Iguana (also known as DZIP1); or bind to one or more of the transcription factors GLI1, GLI2, and GLI3 activated through the pathway By. All these hedgehog pathway proteins are well known human proteins whose sequences are available from UniProtKB/Swiss-Prot and similar data libraries. Because a protein currently has more than one known form in a species due to natural allelic variation between individuals, an inhibitor can bind to and inhibit any or all such known allelic forms, and preferably binds to and inhibits the wild-type, most common or first published allelic form. Exemplary sequences of human SHH, IHH and DHH are assigned UniProtKB/Swiss-Prot accession numbers Q15465, Q14623 and 043323, respectively. Exemplary sequences of other human hedgehog pathway proteins are: PTCH1(Q13635), PTCH2(Q9Y6C5), SMOH(Q99835), DZIP1(Q86YF9), SuFu(Q9UMX1), Gli1(P08151), Gli2(P10070), Gli3(P10071) . The agent can be a protein, such as a mAb, preferably a chimeric, humanized or human mAb, which binds to one or more of the HH protein or PTCH1 (or PTCH2); or can be a small molecule (i.e., has a relatively Low molecular weight, most often compounds below 500 or 600 or 1000 kDa). Exemplary small molecule second agents are cyclopamine, KAAD-cyclopamine (3-keto-N-(aminoethyl-aminocaproyl-dihydrocinnamoyl)cyclopamine), SANT1-4 (Chen et al., Proc. Natl. Acad. Sci. USA2002, 99:14071-14076), CUR61414 (Williams et al., PNAS2003100:84616-4621) and HhAntag-691 (Romer et al., Cancer Cell. 2004; 6:229 -240); JK814Lee, ChemBioChem 2007, 8:1916-1919). Other examples of HH antagonists are described in WO/2004/020599 and Katoh, Cancer Biol Ther. 20054:1050-4 and US Patent No. 7,300,929.

Furthermore, anti-HGF antibodies and fragments of the invention may be used in combination with agonists of PTEN, preferably human PTEN. As used herein, "agonist of PTEN" or "PTEN agonist" means an agent that stimulates the expression of PTEN in a cell or stimulates the activity of PTEN, or which can provide one or more of the functions of PTEN, such as modulating PI3K/Akt/ mTOR path. For example, PTEN can indirectly reduce the activity of mTOR (mammalian target of rapamycin) by down-regulating the activity of Akt. Inhibitors of mTOR directly replicate this specific effect of PTEN (reducing mTOR activity), and so such inhibitors are considered PTEN agonists herein. Such PTEN agonists will replace some, but not necessarily all, functions of the tumor suppressor PTEN in cancer cells with mutated or missing PTEN, and thus can restore the cells to a more normal, less malignant phenotype. Preferred PTEN agonists/mTOR inhibitors for use in the present invention include rapamycin (Rapamune™, sirolimus, ATC code L04AA10, commercially available from Wyeth) and chemical analogs thereof, such as CCI-779 (alternative Sirolimus (temsirolimus, Drug Anatomy, Therapeutics and Chemical Classification System (ATC) code L01XE09, commercially available from Wyeth), RAD-001 (everolimus (everolimus), ATC code L04AA18, commercially available from Novartis ) and AP-2357 (Granville et al., supra). Many PTEN agonists are small molecules (ie, compounds of relatively low molecular weight, most typically below 500 or 600 kDa, or about 1000 kDa in the case of macrolides such as rapamycin). Other agonists include mAbs and zinc finger proteins, or nucleic acids encoding them, engineered to bind to PTEN and activate transcription of PTEN (see eg WO00/00388). Other PTEN agonists are described in US20070280918. Whereas proteins are typically administered parenterally (eg, intravenously), small molecules can be administered parenterally or orally. PTEN and mTOR (also known as FRAP1) are well known human proteins whose sequences are available from UniProtKB/Swiss-Prot and similar data libraries. Because a protein currently has more than one known form in a species due to natural allelic variation between individuals, an inhibitor can bind to and inhibit any or all such known allelic forms, and preferably binds to and inhibits the wild-type, most common or first published allelic form. Exemplary sequences of human PTEN and mTOR (FRAP1) are assigned UniProtKB/Swiss-Prot accession numbers P60484 and P42345. This is intended to be an exemplary and non-exhaustive description of possible combination therapies involving the anti-HGF antibodies and fragments of the invention and another active agent.

The invention also encompasses an in vivo imaging method for detecting the presence of cells expressing HGF, said method comprising administering a diagnostically effective amount of at least one anti-human HGF antibody according to the invention, preferably Ab1, Ab2, Ab7, Ab8, Ab9, Ab10, Ab12, Ab14, Ab19, Ab21, Ab23, Ab24, Ab25 or Ab28. In one embodiment, said administering further comprises administering a radionuclide or fluorophore that facilitates detection of antibodies at the site of the disease expressing HGF. In another embodiment of the invention, the in vivo imaging method is used to detect tumors or metastases expressing HGF, or tumors or metastases expressing HGF-R capable of binding to HGF. In another embodiment, the results of the in vivo imaging method are used to facilitate the design of appropriate treatment regimens, including treatment regimens comprising radiation therapy, chemotherapy, or combinations thereof.

Polynucleotides encoding anti-HGF antibody polypeptides

Antibody Ab1

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:1的重链序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtgcctatgcaatgagctgggtccgccaggctccagagaaggggctggagtggatcgcagtcatttatgttattggtgccactgactacgcgagctgggcgaaaggccgattcaccatctccagaacctcgaccacggtggatctgagaatccccagtccgacaaccgaggacacggccacctatttctgtgccagagtttatgattctgtctggaatcactttaacttgtggggcccgggcaccctggtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggc aaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:11)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:2的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtgcctatgcaatgagctgggtccgccaggctccagagaaggggctggagtggatcgcagtcatttatgttattggtgccactgactacgcgagctgggcgaaaggccgattcaccatctccagaacctcgaccacggtggatctgagaatccccagtccgacaaccgaggacacggccacctatttctgtgccagagtttatgattctgtctggaatcactttaacttgtggggcccgggcaccctggtcaccgtctcgagc(SEQIDNO:12)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:10的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgagg ctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 20).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:21的轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtggggggcacagtcaccatcaagtgccaggccagtcagagcattagcagttggttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctaccaggcatctaaactggcatctggggtcccatcgcggttcaaaggcagtggatctgggacagagttcactctcaccatcagcggcgtggagtgtgccgatgctgccacttactattgtcaacaggcttatagtgttagtaatgttgataatgctttcggcggagggaccgaggtggtggtcaaacgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:31)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:22的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtggggggcacagtcaccatcaagtgccaggccagtcagagcattagcagttggttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctaccaggcatctaaactggcatctggggtcccatcgcggttcaaaggcagtggatctgggacagagttcactctcaccatcagcggcgtggagtgtgccgatgctgccacttactattgtcaacaggcttatagtgttagtaatgttgataatgctttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:32)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:30的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:40)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 14, SEQ ID NO: 16 and SEQ ID NO: 18 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 1 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 2; and/or SEQ ID NO: 34, One or more of the polynucleotide sequences of SEQ ID NO: 36 and SEQ ID NO: 38, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 21 or the variable light chain sequence of SEQ ID NO: 22 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the present invention, the polynucleotide encoding the antibody fragment with binding specificity for HGF comprises or consists of the polynucleosides of SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17 and SEQ ID NO: 19 One or more in the acid sequence, it corresponds to the heavy chain sequence of encoding SEQIDNO:1 or the polynucleotide of framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:2; And/or SEQIDNO: 33. One or more of the polynucleotide sequences of SEQ ID NO: 35, SEQ ID NO: 37 and SEQ ID NO: 39, which correspond to the light chain sequence of SEQ ID NO: 21 or the framework region of the variable light chain sequence of SEQ ID NO: 22 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:11 encoding the heavy chain sequence of SEQ ID NO:1; the polynucleotide SEQ ID NO:12 encoding the variable heavy chain sequence of SEQ ID NO:2; the polynucleotide encoding the light chain sequence of SEQ ID NO:21 Nucleotide SEQIDNO:31; Polynucleotide SEQIDNO:32 encoding the variable light chain sequence of SEQIDNO:22; The complementarity determining region (SEQIDNO:14 of the heavy chain sequence of encoding SEQIDNO:1 or the variable heavy chain sequence of SEQIDNO:2 , SEQIDNO:16 and SEQIDNO:18) polynucleotides; encoding the light chain sequence of SEQIDNO:21 or the complementarity determining region (SEQIDNO:34, SEQIDNO:36 and SEQIDNO:38) of the variable light chain sequence of SEQIDNO:22 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 1 or the framework region (SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17 and SEQ ID NO: 19) of the variable heavy chain sequence of SEQ ID NO: 2; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 21 or the framework region (SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37 and SEQ ID NO: 39) of the variable light chain sequence of SEQ ID NO: 22.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab1, the polynucleotide encoding the full-length Ab1 antibody comprises or consists of the polynucleotide SEQ ID NO:11 encoding the heavy chain sequence of SEQ ID NO:1 and the polynucleotide SEQ ID NO:11 encoding the light chain sequence of SEQ ID NO:21: 31.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl after expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, anti-HGF antibodies such as Ab1 or its Fab fragment can be expressed in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab2

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:41的重链序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtgcctatgcaatgagctgggtccgtcaggctccagggaaggggctggagtgggtcgcagtcatctatgttattggtgccactgactacgcgagcagtgcgaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagagtttatgattctgtctggaatcacttcaacttgtggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactg gctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:51)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:42的可变重链多肽序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtgcctatgcaatgagctgggtccgtcaggctccagggaaggggctggagtgggtcgcagtcatctatgttattggtgccactgactacgcgagcagtgcgaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagagtttatgattctgtctggaatcacttcaacttgtggggccaagggaccctcgtcaccgtctcgagc(SEQIDNO:52)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:50的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgagg ctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 60).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:61的轻链多肽序列的以下多核苷酸序列或者由其组成：gacatccagatgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgccaggccagtcagagcattagcagttggttagcctggtatcagcagaaaccaggaaaagcccctaagctcctgatctatcaggcatctaaactggcatctggagtcccatcaaggttcagcggcagtggatctggaacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttattactgccaacaggcttatagtgttagtaatgttgataatgctttcggcggaggaaccaaggtggaaatcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:71)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:62的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gacatccagatgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgccaggccagtcagagcattagcagttggttagcctggtatcagcagaaaccaggaaaagcccctaagctcctgatctatcaggcatctaaactggcatctggagtcccatcaaggttcagcggcagtggatctggaacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttattactgccaacaggcttatagtgttagtaatgttgataatgctttcggcggaggaaccaaggtggaaatcaaacgt(SEQIDNO:72)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:70的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:80)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO:54, SEQ ID NO:56 and SEQ ID NO:58 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 41 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 42; and/or SEQ ID NO: 74, One or more of the polynucleotide sequences of SEQ ID NO: 76 and SEQ ID NO: 78, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 61 or the variable light chain sequence of SEQ ID NO: 62 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleosides of SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57 and SEQ ID NO: 59 One or more in the acid sequence, it corresponds to the heavy chain sequence of encoding SEQIDNO:41 or the polynucleotide of framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:42; And/or SEQIDNO: 73. One or more of the polynucleotide sequences of SEQ ID NO: 75, SEQ ID NO: 77 and SEQ ID NO: 79, which correspond to the light chain sequence of SEQ ID NO: 61 or the framework region of the variable light chain sequence of SEQ ID NO: 62 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three of the following polynucleotides encoding an antibody fragment ( Including all) or consisting of: the polynucleotide SEQ ID NO:51 encoding the heavy chain sequence of SEQ ID NO:41; the polynucleotide SEQ ID NO:52 encoding the variable heavy chain sequence of SEQ ID NO:42; the light chain sequence encoding SEQ ID NO:61 The polynucleotide of SEQIDNO:71; the polynucleotide of SEQIDNO:72 encoding the variable light chain sequence of SEQIDNO:62; the complementarity determining region (SEQIDNO:72) of the heavy chain sequence of encoding SEQIDNO:41 or the variable heavy chain sequence of SEQIDNO:42 :54, SEQIDNO:56 and SEQIDNO:58) polynucleotides; encoding the light chain sequence of SEQIDNO:61 or the complementarity determining region (SEQIDNO:74, SEQIDNO:76 and SEQIDNO:78) of the variable light chain sequence of SEQIDNO:62 ) polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 41 or the framework region (SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57 and SEQ ID NO: 59) of the variable heavy chain sequence of SEQ ID NO: 42; and A polynucleotide encoding the light chain sequence of SEQ ID NO:61 or the framework region (SEQ ID NO:73, SEQ ID NO:75, SEQ ID NO:77 and SEQ ID NO:79) of the variable light chain sequence of SEQ ID NO:62.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab2, the polynucleotide encoding the full-length Ab2 antibody comprises or consists of the polynucleotide SEQ ID NO:51 encoding the heavy chain sequence of SEQ ID NO:41 and the polynucleotide SEQ ID NO:61 encoding the light chain sequence: 71.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab2 following expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, anti-HGF antibodies such as Ab2 or its Fab fragments can be expressed via Ab2 polynucleotides in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab3

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:81的重链序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccttgacactcacctgcacagtctctggactcaccattagtagctactacatgagctgggtccgccaggctccagggaaggggctggaatggatcggaaccattaatcctggtgctaacacatacttcgcgagctgggcaaaaggccgattcaccatctccagaacctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacatatttctgtgccagagagggggatagtaatgactggggtgtctttgacttgtggggccagggcaccctggtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaa tggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:91)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:82的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccttgacactcacctgcacagtctctggactcaccattagtagctactacatgagctgggtccgccaggctccagggaaggggctggaatggatcggaaccattaatcctggtgctaacacatacttcgcgagctgggcaaaaggccgattcaccatctccagaacctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacatatttctgtgccagagagggggatagtaatgactggggtgtctttgacttgtggggccagggcaccctggtcaccgtctcgagc(SEQIDNO:92)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:90的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgagg ctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 100).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:101的轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtagagatagctgtgggaggcacagtcaccatcaggtgccaggccagtgaggacattgaaagctatttagcctggtatcagcagaaaccagggcagcctcccaaactcctgatctacagggcatccgatctggcatctggggtctcatcgcggttcaaaggcagtggatctgggacagactacactctcaccatcagcggcgtggagtgtgacgatgctgccacttactactgtcaacagggttatactatcgataatgttgataatactttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:111)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:102的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtagagatagctgtgggaggcacagtcaccatcaggtgccaggccagtgaggacattgaaagctatttagcctggtatcagcagaaaccagggcagcctcccaaactcctgatctacagggcatccgatctggcatctggggtctcatcgcggttcaaaggcagtggatctgggacagactacactctcaccatcagcggcgtggagtgtgacgatgctgccacttactactgtcaacagggttatactatcgataatgttgataatactttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:112)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:110的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:120)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 94, SEQ ID NO: 96 and SEQ ID NO: 98 One or more, it corresponds to the polynucleotide encoding the heavy chain sequence of SEQ ID NO:81 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO:82; And/or SEQ ID NO:114, One or more of the polynucleotide sequences of SEQ ID NO: 116 and SEQ ID NO: 118, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 101 or the variable light chain sequence of SEQ ID NO: 102 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97 and SEQ ID NO: 99 One or more in the acid sequence, it corresponds to the heavy chain sequence of encoding SEQIDNO:81 or the polynucleotide of framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:82; And/or SEQIDNO: 113. One or more of the polynucleotide sequences of SEQ ID NO: 115, SEQ ID NO: 117 and SEQ ID NO: 119, which correspond to the light chain sequence of SEQ ID NO: 101 or the framework region of the variable light chain sequence of SEQ ID NO: 102 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:91 encoding the heavy chain sequence of SEQ ID NO:81; the polynucleotide SEQ ID NO:92 encoding the variable heavy chain sequence of SEQ ID NO:82; the polynucleotide encoding the light chain sequence of SEQ ID NO:101 Nucleotide of SEQIDNO:111; polynucleotide SEQIDNO:112 encoding the variable light chain sequence of SEQIDNO:102; the complementarity determining region (SEQIDNO:94) of the heavy chain sequence encoding SEQIDNO:81 or the variable heavy chain sequence of SEQIDNO:82 , SEQIDNO:96 and SEQIDNO:98) polynucleotides; encoding the light chain sequence of SEQIDNO:101 or the complementarity determining region (SEQIDNO:114, SEQIDNO:116 and SEQIDNO:118) of the variable light chain sequence of SEQIDNO:102 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 81 or the framework region (SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97 and SEQ ID NO: 99) of the variable heavy chain sequence of SEQ ID NO: 82; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 101 or the framework region (SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117 and SEQ ID NO: 119) of the variable light chain sequence of SEQ ID NO: 102.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab3, the polynucleotide encoding the full-length Ab3 antibody comprises or consists of the polynucleotide SEQ ID NO:91 encoding the heavy chain sequence of SEQ ID NO:81 and the polynucleotide SEQ ID NO:101 encoding the light chain sequence: 111.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab3 after expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, an anti-HGF antibody such as Ab3 or its Fab fragment can be expressed via Ab3 polynucleotide in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab4

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:121的重链序列的以下多核苷酸序列或者由其组成：cagtcgctggaggagtctgggggacgcttggtccagcctgggacacccctgacactctcctgtacagcctctggactcaccattagtagctactacatgagctgggtccgtcaggctccagggaaggggctggagtgggtcggaaccatcaatcctggtgctaacacatacttcgcgagctctgcaaaaggccgattcaccatctccagatcctcgaccaccctggatcttaagatgaccagcccgacagctgaggacactgctacatattactgtgctagagagggggatagtaatgactggggtgtctttgacttgtggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctga atggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:131)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:122的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcgctggaggagtctgggggacgcttggtccagcctgggacacccctgacactctcctgtacagcctctggactcaccattagtagctactacatgagctgggtccgtcaggctccagggaaggggctggagtgggtcggaaccatcaatcctggtgctaacacatacttcgcgagctctgcaaaaggccgattcaccatctccagatcctcgaccaccctggatcttaagatgaccagcccgacagctgaggacactgctacatattactgtgctagagagggggatagtaatgactggggtgtctttgacttgtggggccaagggaccctcgtcaccgtctcgagc(SEQIDNO:132)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:130的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 140).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:141的轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagtctccagcctccgtggaggcagccgtaggaggcacagtcaccatcaggtgtcaggccagtgaggacattgaaagctacttagcctggtatcagcagaaaccagggcagcctcctaagctcctgatctatagggcatccgatctggcatctggggtctcatcaaggttcaaaggcagtggatctgggacagattacactctcaccatcagcggcctggagcctgaagatgctgcaacttactattgtcaacagggttatactatcgataatgttgacaatactttcggcggaggaaccaaggtggaaatcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:151)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:142的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagtctccagcctccgtggaggcagccgtaggaggcacagtcaccatcaggtgtcaggccagtgaggacattgaaagctacttagcctggtatcagcagaaaccagggcagcctcctaagctcctgatctatagggcatccgatctggcatctggggtctcatcaaggttcaaaggcagtggatctgggacagattacactctcaccatcagcggcctggagcctgaagatgctgcaacttactattgtcaacagggttatactatcgataatgttgacaatactttcggcggaggaaccaaggtggaaatcaaacgt(SEQIDNO:152)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:150的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:160)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 134, SEQ ID NO: 136 and SEQ ID NO: 138 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 121 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 122; and/or SEQ ID NO: 154, One or more of the polynucleotide sequences of SEQ ID NO: 156 and SEQ ID NO: 158, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 141 or the variable light chain sequence of SEQ ID NO: 142 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137 and SEQ ID NO: 139 One or more in the acid sequence, it corresponds to the heavy chain sequence of encoding SEQIDNO:121 or the polynucleotide of framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:122; And/or SEQIDNO: 153. One or more of the polynucleotide sequences of SEQ ID NO: 155, SEQ ID NO: 157 and SEQ ID NO: 159, which correspond to the light chain sequence of SEQ ID NO: 141 or the framework region of the variable light chain sequence of SEQ ID NO: 142 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO: 131 encoding the heavy chain sequence of SEQ ID NO: 121; the polynucleotide SEQ ID NO: 132 encoding the variable heavy chain sequence of SEQ ID NO: 122; the polynucleotide encoding the light chain sequence of SEQ ID NO: 141 Nucleotide of SEQIDNO:151; Polynucleotide SEQIDNO:152 encoding the variable light chain sequence of SEQIDNO:142; The complementarity determining region (SEQIDNO:134 of the heavy chain sequence of encoding SEQIDNO:121 or the variable heavy chain sequence of SEQIDNO:122 , SEQIDNO:136 and SEQIDNO:138) polynucleotides; encoding the light chain sequence of SEQIDNO:141 or the complementarity determining region (SEQIDNO:154, SEQIDNO:156 and SEQIDNO:158) of the variable light chain sequence of SEQIDNO:142 A polynucleotide; a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 121 or the framework region (SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137 and SEQ ID NO: 139) of the variable heavy chain sequence of SEQ ID NO: 122; and encoding SEQ ID NO A polynucleotide of the light chain sequence of SEQ ID NO: 141 or the framework region (SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157 and SEQ ID NO: 159) of the variable light chain sequence of SEQ ID NO: 142.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab4, the polynucleotide encoding the full-length Ab4 antibody comprises or consists of the polynucleotide SEQ ID NO: 131 encoding the heavy chain sequence of SEQ ID NO: 121 and the polynucleotide SEQ ID NO: 141 encoding the light chain sequence: 151.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab4 following expression of the full-length polynucleotide in a suitable host. In another embodiment of the invention, an anti-HGF antibody such as Ab4 or its Fab fragment can be expressed via Ab4 polynucleotide in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab5

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:161的重链序列的以下多核苷酸序列或者由其组成：cagtcgctggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcaataattatgcagtgggctgggtccgccaggctccagggaaggggctggaatggatcggaatcatttaccttagtggtaacacagactacgcgaactgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccaggaaatttgatacgggatatgacatctggggcccaggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagt acaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:171)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:162的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcgctggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcaataattatgcagtgggctgggtccgccaggctccagggaaggggctggaatggatcggaatcatttaccttagtggtaacacagactacgcgaactgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccaggaaatttgatacgggatatgacatctggggcccaggcaccctcgtcaccgtctcgagc(SEQIDNO:172)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:170的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 180).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:181的轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctatggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtacctacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatgatgcatccgatctggcatctggggtctcatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcggcgtggagtgtgacgatgctgccacttactactgtcaacaggattggagtgatagtaatgttgataatgctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:191)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:182的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctatggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtacctacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatgatgcatccgatctggcatctggggtctcatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcggcgtggagtgtgacgatgctgccacttactactgtcaacaggattggagtgatagtaatgttgataatgctttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:192)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:190的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:200)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 174, SEQ ID NO: 176 and SEQ ID NO: 178 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 161 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 162; and/or SEQ ID NO: 194, One or more of the polynucleotide sequences of SEQ ID NO: 196 and SEQ ID NO: 198, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 181 or the variable light chain sequence of SEQ ID NO: 182 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 173, SEQ ID NO: 175, SEQ ID NO: 177 and SEQ ID NO: 179 One or more in the acid sequence, it is corresponding to the heavy chain sequence of encoding SEQIDNO:161 or the polynucleotide of framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:162; And/or SEQIDNO: 193. One or more of the polynucleotide sequences of SEQ ID NO: 195, SEQ ID NO: 197 and SEQ ID NO: 199, which correspond to the light chain sequence of SEQ ID NO: 181 or the framework region of the variable light chain sequence of SEQ ID NO: 182 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO: 171 encoding the heavy chain sequence of SEQ ID NO: 161; the polynucleotide SEQ ID NO: 172 encoding the variable heavy chain sequence of SEQ ID NO: 162; the polynucleotide encoding the light chain sequence of SEQ ID NO: 181 Nucleotide of SEQIDNO:191; Polynucleotide SEQIDNO:192 encoding the variable light chain sequence of SEQIDNO:182; The complementarity determining region (SEQIDNO:174 of the heavy chain sequence of encoding SEQIDNO:161 or the variable heavy chain sequence of SEQIDNO:162 , SEQIDNO:176 and SEQIDNO:178) polynucleotides; encoding the light chain sequence of SEQIDNO:181 or the complementarity determining region (SEQIDNO:194, SEQIDNO:196 and SEQIDNO:198) of the variable light chain sequence of SEQIDNO:182 A polynucleotide; a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 161 or the framework region (SEQ ID NO: 173, SEQ ID NO: 175, SEQ ID NO: 177 and SEQ ID NO: 179) of the variable heavy chain sequence of SEQ ID NO: 162; and encoding SEQ ID NO Polynucleotides of the light chain sequence of: 181 or the framework region (SEQ ID NO: 193, SEQ ID NO: 195, SEQ ID NO: 197 and SEQ ID NO: 199) of the variable light chain sequence of SEQ ID NO: 182.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab5, the polynucleotide encoding the full-length Ab5 antibody comprises or consists of the polynucleotide SEQ ID NO:171 encoding the heavy chain sequence of SEQ ID NO:161 and the polynucleotide SEQ ID NO:181 encoding the light chain sequence: 191.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab5 following expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, anti-HGF antibodies such as Ab5 or its Fab fragments can be expressed in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab6

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:201的重链序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcatgcctgggacacccctgacactcacctgcaccgtctctggattctccctcagtagcaatgcaataagctgggtccgccaggctccagagaaggggctggaatggatcggagtcatttatgttattggtgtcactgactacgcgagctgggcgcaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatccccagtccgacaaccgaggacacggccacctatttctgtgccagagtttatgattctggctggaatcactttaacttgtggggcccgggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatg gcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:211)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:202的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcatgcctgggacacccctgacactcacctgcaccgtctctggattctccctcagtagcaatgcaataagctgggtccgccaggctccagagaaggggctggaatggatcggagtcatttatgttattggtgtcactgactacgcgagctgggcgcaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatccccagtccgacaaccgaggacacggccacctatttctgtgccagagtttatgattctggctggaatcactttaacttgtggggcccgggcaccctcgtcaccgtctcgagc(SEQIDNO:212)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:210的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 220).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:221的轻链多肽序列的以下多核苷酸序列或者由其组成：gctgacattgtgatgacccagactccatcctccgtggaggcagctgtgggaggcacagtcaccatcaagtgccaggccagtgagaacatttataggttattggcctggtatcagcagaaaccagggcagcgtcccaagctcctgatctattctgcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaaactattattatagtagtaggagtagttatgatacatataatgttttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:231)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:222的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gctgacattgtgatgacccagactccatcctccgtggaggcagctgtgggaggcacagtcaccatcaagtgccaggccagtgagaacatttataggttattggcctggtatcagcagaaaccagggcagcgtcccaagctcctgatctattctgcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaaactattattatagtagtaggagtagttatgatacatataatgttttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:232)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:230的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:240)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 214, SEQ ID NO: 216 and SEQ ID NO: 218 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 201 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 202; and/or SEQ ID NO: 234, One or more of the polynucleotide sequences of SEQ ID NO: 236 and SEQ ID NO: 238, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 221 or the variable light chain sequence of SEQ ID NO: 222 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the present invention, the polynucleotide encoding the antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 213, SEQ ID NO: 215, SEQ ID NO: 217 and SEQ ID NO: 219 One or more in the acid sequence, it is corresponding to the heavy chain sequence of encoding SEQIDNO:201 or the polynucleotide of framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:202; And/or SEQIDNO: 233, one or more of the polynucleotide sequences of SEQIDNO:235, SEQIDNO:237 and SEQIDNO:239, which correspond to the light chain sequence of SEQIDNO:221 or the framework region of the variable light chain sequence of SEQIDNO:222 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:211 encoding the heavy chain sequence of SEQ ID NO:201; the polynucleotide SEQ ID NO:212 encoding the variable heavy chain sequence of SEQ ID NO:202; the polynucleotide encoding the light chain sequence of SEQ ID NO:221 Nucleotide SEQIDNO:231; Polynucleotide SEQIDNO:232 encoding the variable light chain sequence of SEQIDNO:222; The complementarity determining region (SEQIDNO:214 of the heavy chain sequence of encoding SEQIDNO:201 or the variable heavy chain sequence of SEQIDNO:202 , SEQIDNO:216 and SEQIDNO:218) polynucleotides; encoding the light chain sequence of SEQIDNO:221 or the complementarity determining region (SEQIDNO:234, SEQIDNO:236 and SEQIDNO:238) of the variable light chain sequence of SEQIDNO:222 A polynucleotide; a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 201 or the framework region (SEQ ID NO: 213, SEQ ID NO: 215, SEQ ID NO: 217 and SEQ ID NO: 219) of the variable heavy chain sequence of SEQ ID NO: 202; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 221 or the framework region (SEQ ID NO: 233, SEQ ID NO: 235, SEQ ID NO: 237 and SEQ ID NO: 239) of the variable light chain sequence of SEQ ID NO: 222.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab6, the polynucleotide encoding the full-length Ab6 antibody comprises or consists of the polynucleotide SEQ ID NO:211 encoding the heavy chain sequence of SEQ ID NO:201 and the polynucleotide SEQ ID NO:221 encoding the light chain sequence: 231.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab6 following expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, an anti-HGF antibody such as Ab6 or its Fab fragment can be expressed via Ab6 polynucleotide in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab7

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:241的重链序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcatgcctgggacacccctgacactcacctgcaccgtctctggattctccctcagtagcaatgcaataagctgggtccgccaggctccagagaaggggctggaatggatcggagtcatttatgttattggtgtcactgactacgcgagctgggcgcaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatccccagtccgacaaccgaggacacggccacctatttctgtgccagagtttatgattctggctggaatcactttaacttgtggggcccagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatg gcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:251)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:242的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcatgcctgggacacccctgacactcacctgcaccgtctctggattctccctcagtagcaatgcaataagctgggtccgccaggctccagagaaggggctggaatggatcggagtcatttatgttattggtgtcactgactacgcgagctgggcgcaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatccccagtccgacaaccgaggacacggccacctatttctgtgccagagtttatgattctggctggaatcactttaacttgtggggcccagggaccctcgtcaccgtctcgagc(SEQIDNO:252)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:250的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 260).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:261的轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagcagttggttagcctggtatcagcagaaaccagggcagcctccaaagctcctgatctacgaagcatccaaactggcatctggggtcccatcgcggttcagtggcagtggatctgggacacagttcactctcaccatcagcggcgtggagtgtgccgatgctgccacttactactgtcaacaggcttatagtgttgccaatgttgataatgctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:271)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:262的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagcagttggttagcctggtatcagcagaaaccagggcagcctccaaagctcctgatctacgaagcatccaaactggcatctggggtcccatcgcggttcagtggcagtggatctgggacacagttcactctcaccatcagcggcgtggagtgtgccgatgctgccacttactactgtcaacaggcttatagtgttgccaatgttgataatgctttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:272)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:270的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:280)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 254, SEQ ID NO: 256 and SEQ ID NO: 258 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 241 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 242; and/or SEQ ID NO: 274, One or more of the polynucleotide sequences of SEQ ID NO: 276 and SEQ ID NO: 278, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 261 or the variable light chain sequence of SEQ ID NO: 262 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 253, SEQ ID NO: 255, SEQ ID NO: 257 and SEQ ID NO: 259 One or more in the acid sequence, it is corresponding to the polynucleotide of the heavy chain sequence of encoding SEQIDNO:241 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:242; And/or SEQIDNO: 273, one or more of the polynucleotide sequences of SEQIDNO:275, SEQIDNO:277 and SEQIDNO:279, which correspond to the light chain sequence of SEQIDNO:261 or the framework region of the variable light chain sequence of SEQIDNO:262 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:251 encoding the heavy chain sequence of SEQ ID NO:241; the polynucleotide SEQ ID NO:252 encoding the variable heavy chain sequence of SEQ ID NO:242; the polynucleotide encoding the light chain sequence of SEQ ID NO:261 Nucleotide SEQIDNO:271; Polynucleotide SEQIDNO:272 encoding the variable light chain sequence of SEQIDNO:262; The complementarity determining region (SEQIDNO:254 of the heavy chain sequence of encoding SEQIDNO:241 or the variable heavy chain sequence of SEQIDNO:242 , SEQIDNO:256 and SEQIDNO:258) polynucleotides; encoding the light chain sequence of SEQIDNO:261 or the complementarity determining region (SEQIDNO:274, SEQIDNO:276 and SEQIDNO:278) of the variable light chain sequence of SEQIDNO:262 A polynucleotide; a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 241 or the framework region (SEQ ID NO: 253, SEQ ID NO: 255, SEQ ID NO: 257 and SEQ ID NO: 259) of the variable heavy chain sequence of SEQ ID NO: 242; and encoding SEQ ID NO A polynucleotide of the light chain sequence of 261 or the framework region (SEQ ID NO: 273, SEQ ID NO: 275, SEQ ID NO: 277 and SEQ ID NO: 279) of the variable light chain sequence of SEQ ID NO: 262.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab7, the polynucleotide encoding the full-length Ab7 antibody comprises or consists of the polynucleotide SEQ ID NO:251 encoding the heavy chain sequence of SEQ ID NO:241 and the polynucleotide SEQ ID NO:261 encoding the light chain sequence: 271.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab7 following expression of the full-length polynucleotide in a suitable host. In another embodiment of the invention, an anti-HGF antibody such as Ab7 or its Fab fragment can be expressed via Ab7 polynucleotide in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab8

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:281的重链序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcaatgcaataagctgggtccgtcaggctccagggaaggggctggagtgggtcggagtcatttatgttattggtgtcactgactacgcgagctctgcgcaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagagtttatgattctggctggaatcactttaacttgtggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggact ggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:291)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:282的可变重链多肽序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcaatgcaataagctgggtccgtcaggctccagggaaggggctggagtgggtcggagtcatttatgttattggtgtcactgactacgcgagctctgcgcaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagagtttatgattctggctggaatcactttaacttgtggggccaagggaccctcgtcaccgtctcgagc(SEQIDNO:292)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:290的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 300).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:301的轻链多肽序列的以下多核苷酸序列或者由其组成：gacatccagatgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgccaggccagtcagagcattagcagttggttagcctggtatcagcagaaaccaggaaaagcccctaagctcctgatctatgaagcatccaaactggcatctggagtcccatcaaggttcagcggcagtggatctggaacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttattactgccaacaggcttatagtgttgccaatgttgataatgctttcggcggaggaaccaaggtggaaatcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:311)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:302的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gacatccagatgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgccaggccagtcagagcattagcagttggttagcctggtatcagcagaaaccaggaaaagcccctaagctcctgatctatgaagcatccaaactggcatctggagtcccatcaaggttcagcggcagtggatctggaacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttattactgccaacaggcttatagtgttgccaatgttgataatgctttcggcggaggaaccaaggtggaaatcaaacgt(SEQIDNO:312)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:310的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:320)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 294, SEQ ID NO: 296 and SEQ ID NO: 298 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 281 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 282; and/or SEQ ID NO: 314, One or more of the polynucleotide sequences of SEQ ID NO: 316 and SEQ ID NO: 318, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 301 or the variable light chain sequence of SEQ ID NO: 302 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 293, SEQ ID NO: 295, SEQ ID NO: 297 and SEQ ID NO: 299 One or more in the acid sequence, it corresponds to the polynucleotide of the heavy chain sequence of encoding SEQIDNO:281 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:282; And/or SEQIDNO: 313, one or more of the polynucleotide sequences of SEQIDNO:315, SEQIDNO:317 and SEQIDNO:319, which correspond to the light chain sequence of SEQIDNO:301 or the framework region of the variable light chain sequence of SEQIDNO:302 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:291 encoding the heavy chain sequence of SEQ ID NO:281; the polynucleotide SEQ ID NO:292 encoding the variable heavy chain sequence of SEQ ID NO:282; the polynucleotide encoding the light chain sequence of SEQ ID NO:301 Nucleotide SEQIDNO:311; Polynucleotide SEQIDNO:312 encoding the variable light chain sequence of SEQIDNO:302; The complementarity determining region (SEQIDNO:294 of the heavy chain sequence of encoding SEQIDNO:281 or the variable heavy chain sequence of SEQIDNO:282 , the polynucleotides of SEQ ID NO: 296 and SEQ ID NO: 298; the polynucleotides encoding the light chain sequence of SEQ ID NO: 301 or the complementarity determining region (SEQ ID NO: 314, SEQ ID NO: 316 and SEQ ID NO: 318) of the variable light chain sequence of SEQ ID NO: 302 Nucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 281 or the framework region (SEQ ID NO: 293, SEQ ID NO: 295, SEQ ID NO: 297 and SEQ ID NO: 299) of the variable heavy chain sequence of SEQ ID NO: 282; and encoding SEQ ID NO: A polynucleotide of the light chain sequence of 301 or the framework region (SEQ ID NO: 313, SEQ ID NO: 315, SEQ ID NO: 317 and SEQ ID NO: 319) of the variable light chain sequence of SEQ ID NO: 302.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab8, the polynucleotide encoding the full-length Ab8 antibody comprises or consists of the polynucleotide SEQ ID NO:291 encoding the heavy chain sequence of SEQ ID NO:281 and the polynucleotide SEQ ID NO:291 encoding the light chain sequence of SEQ ID NO:301: 311.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab8 following expression of the full-length polynucleotide in a suitable host. In another embodiment of the invention, an anti-HGF antibody such as Ab8 or its Fab fragment can be expressed in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab9

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:321的重链序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtgtctggaatcgacctcaatagcaatggaatgagctgggtccgccaggctccaggggaggggctggaatggatcggagccagtagtattgatgggaccacatactacaccaattgggcgaagggccgattcaccatctccaaaacctcgtcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtaccagaggggagtatgctggtgttgttggttcgaactactttgacttgtggggccagggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccagg actggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:331)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:322的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtgtctggaatcgacctcaatagcaatggaatgagctgggtccgccaggctccaggggaggggctggaatggatcggagccagtagtattgatgggaccacatactacaccaattgggcgaagggccgattcaccatctccaaaacctcgtcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtaccagaggggagtatgctggtgttgttggttcgaactactttgacttgtggggccagggcaccctcgtcaccgtctcgagc(SEQIDNO:332)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:330的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 340).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:341的轻链多肽序列的以下多核苷酸序列或者由其组成：caagtgctgacccagactccaccctccgtgtctgcagttgtgggaggcacagtcaccatcaattgccagtccagtcagaggatttatagtaattggttatcttggtatcagcagaaaccagggcagactcccaagcccctgatctatgctgcatccagcctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcgacctggagtgtgacgatgctgccagttactactgtgcaggctattatagtggtcatatttattctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:351)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:342的可变轻链多肽序列的以下多核苷酸序列或者由其组成：caagtgctgacccagactccaccctccgtgtctgcagttgtgggaggcacagtcaccatcaattgccagtccagtcagaggatttatagtaattggttatcttggtatcagcagaaaccagggcagactcccaagcccctgatctatgctgcatccagcctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcgacctggagtgtgacgatgctgccagttactactgtgcaggctattatagtggtcatatttattctttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:352)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:350的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:360)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 334, SEQ ID NO: 336 and SEQ ID NO: 338 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 321 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 322; and/or SEQ ID NO: 354, One or more of the polynucleotide sequences of SEQ ID NO: 356 and SEQ ID NO: 358, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 341 or the variable light chain sequence of SEQ ID NO: 342 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 333, SEQ ID NO: 335, SEQ ID NO: 337 and SEQ ID NO: 339 One or more in the acid sequence, it is corresponding to the polynucleotide of the heavy chain sequence of encoding SEQIDNO:321 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:322; And/or SEQIDNO: 353, one or more of the polynucleotide sequences of SEQIDNO:355, SEQIDNO:357 and SEQIDNO:359, which correspond to the light chain sequence of SEQIDNO:341 or the framework region of the variable light chain sequence of SEQIDNO:342 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:331 encoding the heavy chain sequence of SEQ ID NO:321; the polynucleotide SEQ ID NO:332 encoding the variable heavy chain sequence of SEQ ID NO:322; the polynucleotide encoding the light chain sequence of SEQ ID NO:341 Nucleotide SEQIDNO:351; Polynucleotide SEQIDNO:352 encoding the variable light chain sequence of SEQIDNO:342; The complementarity determining region (SEQIDNO:334 of the heavy chain sequence of encoding SEQIDNO:321 or the variable heavy chain sequence of SEQIDNO:322 , SEQIDNO:336 and SEQIDNO:338) polynucleotides; encoding the light chain sequence of SEQIDNO:341 or the complementarity determining region (SEQIDNO:354, SEQIDNO:356 and SEQIDNO:358) of the variable light chain sequence of SEQIDNO:342 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 321 or the framework region (SEQ ID NO: 333, SEQ ID NO: 335, SEQ ID NO: 337 and SEQ ID NO: 339) of the variable heavy chain sequence of SEQ ID NO: 322; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 341 or the framework region (SEQ ID NO: 353, SEQ ID NO: 355, SEQ ID NO: 357 and SEQ ID NO: 359) of the variable light chain sequence of SEQ ID NO: 342.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab9, the polynucleotide encoding the full-length Ab9 antibody comprises or consists of the polynucleotide SEQ ID NO: 331 encoding the heavy chain sequence of SEQ ID NO: 321 and the polynucleotide SEQ ID NO: 341 encoding the light chain sequence: 351.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab9 following expression of the full-length polynucleotide in a suitable host. In another embodiment of the invention, an anti-HGF antibody such as Ab9 or its Fab fragment can be expressed in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab10

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:361的重链序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcaatggaatgagctgggtccgtcaggctccagggaaggggctggagtgggtcggagccagtagtattgatgggaccacatactacaccaattctgcgaagggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagaggggagtatgctggtgttgttggttcgaactactttgacttgtggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgc accaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:371)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:362的可变重链多肽序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcaatggaatgagctgggtccgtcaggctccagggaaggggctggagtgggtcggagccagtagtattgatgggaccacatactacaccaattctgcgaagggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagaggggagtatgctggtgttgttggttcgaactactttgacttgtggggccaagggaccctcgtcaccgtctcgagc(SEQIDNO:372)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:370的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 380).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:381的轻链多肽序列的以下多核苷酸序列或者由其组成：gacatccagatgacccagtctccatcttccgtgtctgcatctgtaggagacagagtcaccatcacttgtcagtccagtcagaggatctatagtaattggttatcttggtatcagcagaaaccagggaaagcccctaagctcctgatctatgctgcatccagcctggcatctggggtcccatcaaggttcagcggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctgaagattttgcaacttactattgtgcaggctactatagtggtcatatctattctttcggcggaggaaccaaggtggaaatcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:391)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:382的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gacatccagatgacccagtctccatcttccgtgtctgcatctgtaggagacagagtcaccatcacttgtcagtccagtcagaggatctatagtaattggttatcttggtatcagcagaaaccagggaaagcccctaagctcctgatctatgctgcatccagcctggcatctggggtcccatcaaggttcagcggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctgaagattttgcaacttactattgtgcaggctactatagtggtcatatctattctttcggcggaggaaccaaggtggaaatcaaacgt(SEQIDNO:392)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:390的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:400)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 374, SEQ ID NO: 376 and SEQ ID NO: 378 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 361 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 362; and/or SEQ ID NO: 394, One or more of the polynucleotide sequences of SEQ ID NO: 396 and SEQ ID NO: 398, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 381 or the variable light chain sequence of SEQ ID NO: 382 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 373, SEQ ID NO: 375, SEQ ID NO: 377 and SEQ ID NO: 379 One or more of acid sequences corresponding to the polynucleotide encoding the heavy chain sequence of SEQ ID NO: 361 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 362; and/or SEQ ID NO: 393, one or more of the polynucleotide sequences of SEQIDNO:395, SEQIDNO:397, and SEQIDNO:399, which correspond to the light chain sequence of SEQIDNO:381 or the framework region of the variable light chain sequence of SEQIDNO:382 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:371 encoding the heavy chain sequence of SEQ ID NO:361; the polynucleotide SEQ ID NO:372 encoding the variable heavy chain sequence of SEQ ID NO:362; the polynucleotide encoding the light chain sequence of SEQ ID NO:381 Nucleotide SEQIDNO:391; Polynucleotide SEQIDNO:392 encoding the variable light chain sequence of SEQIDNO:382; The complementarity determining region (SEQIDNO:374 of the heavy chain sequence of encoding SEQIDNO:361 or the variable heavy chain sequence of SEQIDNO:362 , SEQIDNO:376 and SEQIDNO:378) polynucleotides; encoding the light chain sequence of SEQIDNO:381 or the complementarity determining region (SEQIDNO:394, SEQIDNO:396 and SEQIDNO:398) of the variable light chain sequence of SEQIDNO:382 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 361 or the framework region (SEQ ID NO: 373, SEQ ID NO: 375, SEQ ID NO: 377 and SEQ ID NO: 379) of the variable heavy chain sequence of SEQ ID NO: 362; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 381 or the framework region (SEQ ID NO: 393, SEQ ID NO: 395, SEQ ID NO: 397 and SEQ ID NO: 399) of the variable light chain sequence of SEQ ID NO: 382.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab10, the polynucleotide encoding the full-length Ab10 antibody comprises or consists of the polynucleotide SEQ ID NO:371 encoding the heavy chain sequence of SEQ ID NO:361 and the polynucleotide SEQ ID NO:381 encoding the light chain sequence: 391.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of AblO after expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, anti-HGF antibodies such as Ab10 or its Fab fragments can be expressed in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab11

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:401的重链序列的以下多核苷酸序列或者由其组成：cagtcaatggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtgactatgcgttgagctgggtccgccaggctccagggaaggggctggaatggatcggaatgattagtagtggtgacaacacatactacgcgagctgggcgaaaggccgattcaccatctccaaagcctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagagataaagatgctagtagtggtggttatttggtccttgacctattggatgtccccgacggcatggacctctggggcccaggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtca gcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:411)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:402的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcaatggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtgactatgcgttgagctgggtccgccaggctccagggaaggggctggaatggatcggaatgattagtagtggtgacaacacatactacgcgagctgggcgaaaggccgattcaccatctccaaagcctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagagataaagatgctagtagtggtggttatttggtccttgacctattggatgtccccgacggcatggacctctggggcccaggcaccctcgtcaccgtctcgagc(SEQIDNO:412)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:410的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 420).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:421的轻链多肽序列的以下多核苷酸序列或者由其组成：gccgtgctgacccagacaccatcgcccgtgtctgcagctgtgggaggcacagtcaccatcaagtgccagtccagtcagagtgtttataataacaacctcttatcctggtatcagcagaaaccagggcagcctcccaagctcctgatctggggtgcatcctatctgccatctggggtcccagataggttcagcggcagtggatctgggacacagttcactctcaccatcagcggcgtgcagtgtgacgatgctgccacttactactgtctaggcggttatgatggtgatgctgatacatataatactttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:431)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:422的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gccgtgctgacccagacaccatcgcccgtgtctgcagctgtgggaggcacagtcaccatcaagtgccagtccagtcagagtgtttataataacaacctcttatcctggtatcagcagaaaccagggcagcctcccaagctcctgatctggggtgcatcctatctgccatctggggtcccagataggttcagcggcagtggatctgggacacagttcactctcaccatcagcggcgtgcagtgtgacgatgctgccacttactactgtctaggcggttatgatggtgatgctgatacatataatactttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:432)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:430的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:440)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 414, SEQ ID NO: 416 and SEQ ID NO: 418 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 401 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 402; and/or SEQ ID NO: 434, One or more of the polynucleotide sequences of SEQ ID NO: 436 and SEQ ID NO: 438, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 421 or the variable light chain sequence of SEQ ID NO: 422 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 413, SEQ ID NO: 415, SEQ ID NO: 417 and SEQ ID NO: 419 One or more in acid sequence, it is corresponding to the polynucleotide of the heavy chain sequence of encoding SEQIDNO:401 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:402; And/or SEQIDNO: 433, one or more of the polynucleotide sequences of SEQIDNO:435, SEQIDNO:437 and SEQIDNO:439, which correspond to the light chain sequence of SEQIDNO:421 or the framework region of the variable light chain sequence of SEQIDNO:422 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:411 encoding the heavy chain sequence of SEQ ID NO:401; the polynucleotide SEQ ID NO:412 encoding the variable heavy chain sequence of SEQ ID NO:402; the polynucleotide encoding the light chain sequence of SEQ ID NO:421 Nucleotide SEQIDNO:431; Polynucleotide SEQIDNO:432 encoding the variable light chain sequence of SEQIDNO:422; The complementarity determining region (SEQIDNO:414 of the heavy chain sequence of encoding SEQIDNO:401 or the variable heavy chain sequence of SEQIDNO:402 , SEQIDNO:416 and SEQIDNO:418) polynucleotides; encoding the light chain sequence of SEQIDNO:421 or the complementarity determining region (SEQIDNO:434, SEQIDNO:436 and SEQIDNO:438) of the variable light chain sequence of SEQIDNO:422 A polynucleotide; a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 401 or the framework region (SEQ ID NO: 413, SEQ ID NO: 415, SEQ ID NO: 417 and SEQ ID NO: 419) of the variable heavy chain sequence of SEQ ID NO: 402; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 421 or the framework region (SEQ ID NO: 433, SEQ ID NO: 435, SEQ ID NO: 437 and SEQ ID NO: 439) of the variable light chain sequence of SEQ ID NO: 422.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab11, the polynucleotide encoding the full-length Ab11 antibody comprises or consists of the polynucleotide SEQ ID NO:411 encoding the heavy chain sequence of SEQ ID NO:401 and the polynucleotide SEQ ID NO:421 encoding the light chain sequence: 431.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab11 following expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, an anti-HGF antibody such as Ab11 or its Fab fragment can be expressed via Ab11 polynucleotide in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab12

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:441的重链序列的以下多核苷酸序列或者由其组成：cagtcgctggaggagtccgggggtcgcctggtaacgcctggaggatccctgacactcacctgcacagtctctggaatcgacctcagtagcaatgcaataagctgggtccgccaggctccagagaaggggctggagtggatcgcagtcatttatgttgttggtgccactgactacgcgagctgggcgaaaggccgattcaccatctccagaacctcgaccacggtggatctgaaaatgaccagtctgacaaccgaggacacggccacctatttctgtgccagagtttatgattctggctggaatcactttaacttgtggggcccaggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatg gcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:451)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:442的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcgctggaggagtccgggggtcgcctggtaacgcctggaggatccctgacactcacctgcacagtctctggaatcgacctcagtagcaatgcaataagctgggtccgccaggctccagagaaggggctggagtggatcgcagtcatttatgttgttggtgccactgactacgcgagctgggcgaaaggccgattcaccatctccagaacctcgaccacggtggatctgaaaatgaccagtctgacaaccgaggacacggccacctatttctgtgccagagtttatgattctggctggaatcactttaacttgtggggcccaggcaccctcgtcaccgtctcgagc(SEQIDNO:452)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:450的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 460).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:461的轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggtcagtcagagcattagtagttggttatcctggtatcagaagaaaccagggcagcgtcccaagctcctgatctacagggcatccactctggcatctggggtctcatcgcggttcaaaggcagtggatctgggacagagttcactctcaccatcagcggcgtggagtgtgccgatgctgccacttactactgtcaacaggcttatagtgttagtaatgttgataatgctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:471)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:462的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggtcagtcagagcattagtagttggttatcctggtatcagaagaaaccagggcagcgtcccaagctcctgatctacagggcatccactctggcatctggggtctcatcgcggttcaaaggcagtggatctgggacagagttcactctcaccatcagcggcgtggagtgtgccgatgctgccacttactactgtcaacaggcttatagtgttagtaatgttgataatgctttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:472)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:470的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:480)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 454, SEQ ID NO: 456 and SEQ ID NO: 458 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 441 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 442; and/or SEQ ID NO: 474, One or more of the polynucleotide sequences of SEQ ID NO: 476 and SEQ ID NO: 478, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 461 or the variable light chain sequence of SEQ ID NO: 462 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 453, SEQ ID NO: 455, SEQ ID NO: 457 and SEQ ID NO: 459 One or more in the acid sequence, it corresponds to the polynucleotide of the heavy chain sequence of encoding SEQIDNO:441 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:442; And/or SEQIDNO: 473, one or more of the polynucleotide sequences of SEQIDNO:475, SEQIDNO:477 and SEQIDNO:479, which correspond to the light chain sequence of SEQIDNO:461 or the framework region of the variable light chain sequence of SEQIDNO:462 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:451 encoding the heavy chain sequence of SEQ ID NO:441; the polynucleotide SEQ ID NO:452 encoding the variable heavy chain sequence of SEQ ID NO:442; the polynucleotide encoding the light chain sequence of SEQ ID NO:461 Nucleotide SEQIDNO:471; polynucleotide SEQIDNO:472 encoding the variable light chain sequence of SEQIDNO:462; the complementarity determining region (SEQIDNO:454) of the heavy chain sequence encoding SEQIDNO:441 or the variable heavy chain sequence of SEQIDNO:442 , SEQIDNO:456 and SEQIDNO:458) polynucleotides; encoding the light chain sequence of SEQIDNO:461 or the complementarity determining region (SEQIDNO:474, SEQIDNO:476 and SEQIDNO:478) of the variable light chain sequence of SEQIDNO:462 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 441 or the framework region (SEQ ID NO: 453, SEQ ID NO: 455, SEQ ID NO: 457 and SEQ ID NO: 459) of the variable heavy chain sequence of SEQ ID NO: 442; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 461 or the framework region (SEQ ID NO: 473, SEQ ID NO: 475, SEQ ID NO: 477 and SEQ ID NO: 479) of the variable light chain sequence of SEQ ID NO: 462.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab12, the polynucleotide encoding the full-length Ab12 antibody comprises or consists of the polynucleotide SEQ ID NO:451 encoding the heavy chain sequence of SEQ ID NO:441 and the polynucleotide SEQ ID NO:461 encoding the light chain sequence: 471.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl2 after expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, anti-HGF antibodies such as Ab12 or its Fab fragments can be expressed in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab13

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:481的重链序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcaatgcaataagctgggtccgtcaggctccagggaaggggctggagtgggtcgcagtcatctatgttgttggtgccactgactacgcgagcagtgcgaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagagtttatgattctggctggaatcactttaacttgtggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggact ggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:491)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:482的可变重链多肽序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcaatgcaataagctgggtccgtcaggctccagggaaggggctggagtgggtcgcagtcatctatgttgttggtgccactgactacgcgagcagtgcgaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagagtttatgattctggctggaatcactttaacttgtggggccaagggaccctcgtcaccgtctcgagc(SEQIDNO:492)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:490的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 500).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:501的轻链多肽序列的以下多核苷酸序列或者由其组成：gactatcagatgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgccaggtcagtcagagcattagtagttggttatcctggtatcagcagaaaccaggaaaagcccctaagctcctgatctatagggcatccactctggcatctggagtcccatcaaggttcagcggcagtggatctggaacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttattactgccaacaggcttatagtgttagtaatgttgataatgctttcggcggaggaaccaaggtggaaatcaaacgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:511)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:502的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gactatcagatgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgccaggtcagtcagagcattagtagttggttatcctggtatcagcagaaaccaggaaaagcccctaagctcctgatctatagggcatccactctggcatctggagtcccatcaaggttcagcggcagtggatctggaacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttattactgccaacaggcttatagtgttagtaatgttgataatgctttcggcggaggaaccaaggtggaaatcaaacgt(SEQIDNO:512)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:510的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:520)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 494, SEQ ID NO: 496 and SEQ ID NO: 498 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 481 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 482; and/or SEQ ID NO: 514, One or more of the polynucleotide sequences of SEQ ID NO: 516 and SEQ ID NO: 518, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 501 or the variable light chain sequence of SEQ ID NO: 502 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 493, SEQ ID NO: 495, SEQ ID NO: 497 and SEQ ID NO: 499 One or more in the acid sequence, it corresponds to the heavy chain sequence of encoding SEQIDNO:481 or the polynucleotide of framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:482; And/or SEQIDNO: 513, one or more of the polynucleotide sequences of SEQIDNO:515, SEQIDNO:517 and SEQIDNO:519, which correspond to the light chain sequence of SEQIDNO:501 or the framework region of the variable light chain sequence of SEQIDNO:502 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:491 encoding the heavy chain sequence of SEQ ID NO:481; the polynucleotide SEQ ID NO:492 encoding the variable heavy chain sequence of SEQ ID NO:482; the polynucleotide encoding the light chain sequence of SEQ ID NO:501 Nucleotide SEQIDNO:511; polynucleotide SEQIDNO:512 encoding the variable light chain sequence of SEQIDNO:502; the complementarity determining region (SEQIDNO:494) of the heavy chain sequence encoding SEQIDNO:481 or the variable heavy chain sequence of SEQIDNO:482 , SEQIDNO:496 and SEQIDNO:498) polynucleotides; encoding the light chain sequence of SEQIDNO:501 or the complementarity determining region (SEQIDNO:514, SEQIDNO:516 and SEQIDNO:518) of the variable light chain sequence of SEQIDNO:502 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 481 or the framework region (SEQ ID NO: 493, SEQ ID NO: 495, SEQ ID NO: 497 and SEQ ID NO: 499) of the variable heavy chain sequence of SEQ ID NO: 482; and encoding SEQ ID NO Polynucleotides of the light chain sequence of: 501 or the framework region (SEQ ID NO: 513, SEQ ID NO: 515, SEQ ID NO: 517 and SEQ ID NO: 519) of the variable light chain sequence of SEQ ID NO: 502.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab13, the polynucleotide encoding the full-length Ab13 antibody comprises or consists of the polynucleotide SEQ ID NO:491 encoding the heavy chain sequence of SEQ ID NO:481 and the polynucleotide SEQ ID NO:501 encoding the light chain sequence: 511.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl3 after expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, anti-HGF antibodies such as Ab13 or its Fab fragments can be expressed in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab14

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:521的重链序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtaactacgcaatgacctgggtccgccaggctccagggaaggggctggaatggatcggagtcattagttttggtggtaacacatactacgcgaactgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagatgggatgctgaaaacaatgagattcttaacttgtggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatg gcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:531)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:522的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtaactacgcaatgacctgggtccgccaggctccagggaaggggctggaatggatcggagtcattagttttggtggtaacacatactacgcgaactgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagatgggatgctgaaaacaatgagattcttaacttgtggggccaagggaccctcgtcaccgtctcgagc(SEQIDNO:532)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:530的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 540).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:541的轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtgagagcattgaaagctatttagcctggtatcagcagaaatcagggcagcctcccaagctcctgatctacagggcttccactctggcatctggggtctcatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcggcgtggaatgtgccgatgctgccacttactactgtcaacagggtgatgcttggagtaatgttgataatgttttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:551)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:542的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtgagagcattgaaagctatttagcctggtatcagcagaaatcagggcagcctcccaagctcctgatctacagggcttccactctggcatctggggtctcatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcggcgtggaatgtgccgatgctgccacttactactgtcaacagggtgatgcttggagtaatgttgataatgttttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:552)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:550的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:560)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO:534, SEQ ID NO:536 and SEQ ID NO:538 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 521 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 522; and/or SEQ ID NO: 554, One or more of the polynucleotide sequences of SEQ ID NO: 556 and SEQ ID NO: 558, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 541 or the variable light chain sequence of SEQ ID NO: 542 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 533, SEQ ID NO: 535, SEQ ID NO: 537 and SEQ ID NO: 539 One or more in the acid sequence, it is corresponding to the polynucleotide of the heavy chain sequence of encoding SEQIDNO:521 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:522; And/or SEQIDNO: 553, one or more of the polynucleotide sequences of SEQIDNO:555, SEQIDNO:557 and SEQIDNO:559, which correspond to the light chain sequence of SEQIDNO:541 or the framework region of the variable light chain sequence of SEQIDNO:542 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:531 encoding the heavy chain sequence of SEQ ID NO:521; the polynucleotide SEQ ID NO:532 encoding the variable heavy chain sequence of SEQ ID NO:522; the polynucleotide encoding the light chain sequence of SEQ ID NO:541 Nucleotide SEQIDNO:551; Polynucleotide SEQIDNO:552 encoding the variable light chain sequence of SEQIDNO:542; The complementarity determining region (SEQIDNO:534 of the heavy chain sequence of encoding SEQIDNO:521 or the variable heavy chain sequence of SEQIDNO:522 , SEQIDNO:536 and SEQIDNO:538) polynucleotides; encoding the light chain sequence of SEQIDNO:541 or the complementarity determining region (SEQIDNO:554, SEQIDNO:556 and SEQIDNO:558) of the variable light chain sequence of SEQIDNO:542 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 521 or the framework region (SEQ ID NO: 533, SEQ ID NO: 535, SEQ ID NO: 537 and SEQ ID NO: 539) of the variable heavy chain sequence of SEQ ID NO: 522; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 541 or the framework region (SEQ ID NO: 553, SEQ ID NO: 555, SEQ ID NO: 557 and SEQ ID NO: 559) of the variable light chain sequence of SEQ ID NO: 542.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab14, the polynucleotide encoding the full-length Ab14 antibody comprises or consists of the polynucleotide SEQ ID NO:531 encoding the heavy chain sequence of SEQ ID NO:521 and the polynucleotide SEQ ID NO:541 encoding the light chain sequence: 551.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl4 after expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, anti-HGF antibodies such as Ab14 or its Fab fragments can be expressed in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab15

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:561的重链序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtaactacgcaatgacctgggtccgccaggctccagggaaggggctggaatggatcggagtcattagttttggtggtaacacatactacgcgaactgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagatgggatgctgaaaacaatgagattcttaacttgtggggcccagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatg gcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:571)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:562的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtaactacgcaatgacctgggtccgccaggctccagggaaggggctggaatggatcggagtcattagttttggtggtaacacatactacgcgaactgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagatgggatgctgaaaacaatgagattcttaacttgtggggcccagggaccctcgtcaccgtctcgagc(SEQIDNO:572)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:570的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 580).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:581的轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtgagagcattagtagctacttagcctggtatcagcagaaatcagggcagcctcccaagctcctgatctacagggcttccactctggcatctggggtctcatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcggcgtggagtgtgccgatgctgccacttactactgtcaacagggtgatgcttggagtaatgttgataatgttttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:591)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:582的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtgagagcattagtagctacttagcctggtatcagcagaaatcagggcagcctcccaagctcctgatctacagggcttccactctggcatctggggtctcatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcggcgtggagtgtgccgatgctgccacttactactgtcaacagggtgatgcttggagtaatgttgataatgttttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:592)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:590的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:600)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO:574, SEQ ID NO:576 and SEQ ID NO:578 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 561 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 562; and/or SEQ ID NO: 594, One or more of the polynucleotide sequences of SEQ ID NO: 596 and SEQ ID NO: 598, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 581 or the variable light chain sequence of SEQ ID NO: 582 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO:573, SEQ ID NO:575, SEQ ID NO:577 and SEQ ID NO:579 One or more of the acid sequences corresponding to the polynucleotide encoding the heavy chain sequence of SEQ ID NO: 561 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 562; and/or SEQ ID NO: 593, one or more of the polynucleotide sequences of SEQIDNO:595, SEQIDNO:597 and SEQIDNO:599, which correspond to the light chain sequence of SEQIDNO:581 or the framework region of the variable light chain sequence of SEQIDNO:582 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:571 encoding the heavy chain sequence of SEQ ID NO:561; the polynucleotide SEQ ID NO:572 encoding the variable heavy chain sequence of SEQ ID NO:562; the polynucleotide encoding the light chain sequence of SEQ ID NO:581 Nucleotide SEQIDNO:591; Polynucleotide SEQIDNO:592 encoding the variable light chain sequence of SEQIDNO:582; The complementarity determining region (SEQIDNO:574 of the heavy chain sequence of encoding SEQIDNO:561 or the variable heavy chain sequence of SEQIDNO:562 , SEQIDNO:576 and SEQIDNO:578) polynucleotides; encoding the light chain sequence of SEQIDNO:581 or the complementarity determining region (SEQIDNO:594, SEQIDNO:596 and SEQIDNO:598) of the variable light chain sequence of SEQIDNO:582 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 561 or the framework region (SEQ ID NO: 573, SEQ ID NO: 575, SEQ ID NO: 577 and SEQ ID NO: 579) of the variable heavy chain sequence of SEQ ID NO: 562; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 581 or the framework region (SEQ ID NO: 593, SEQ ID NO: 595, SEQ ID NO: 597 and SEQ ID NO: 599) of the variable light chain sequence of SEQ ID NO: 582.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab15, the polynucleotide encoding the full-length Ab15 antibody comprises or consists of the polynucleotide SEQ ID NO:571 encoding the heavy chain sequence of SEQ ID NO:561 and the polynucleotide SEQ ID NO:581 encoding the light chain sequence: 591.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl5 after expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, anti-HGF antibodies such as Ab15 or its Fab fragments can be expressed via Ab15 polynucleotides in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab16

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:601的重链序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggaatcgacctcagtaattatgcaatgggctgggtccgccaggctccagggaaggggctggaatacatcggaatgattggtgttaatggtagggcatggtacgcgacttgggcgaaaggccgattcaccatctccaagacctcgcccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagattgattgacgagcgttcaacttatagttatgtttttgacttgtggggccaaggcaccctggtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggact ggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:611)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:602的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggaatcgacctcagtaattatgcaatgggctgggtccgccaggctccagggaaggggctggaatacatcggaatgattggtgttaatggtagggcatggtacgcgacttgggcgaaaggccgattcaccatctccaagacctcgcccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagattgattgacgagcgttcaacttatagttatgtttttgacttgtggggccaaggcaccctggtcaccgtctcgagc(SEQIDNO:612)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:610的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 620).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:621的轻链多肽序列的以下多核苷酸序列或者由其组成：caagtgctgacccagactccatcccctgtgtctgcagctgtgggaggcacagtcaccatcaactgccagggcagtcagagtctttataataacaacgccttttcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatgatgcttccactctggcgtctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagtggcgtgcagtgtgcagatgctgccacttactactgtcaaggcgaatttagttgtggtgatgttgattgtattgctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:631)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:622的可变轻链多肽序列的以下多核苷酸序列或者由其组成：caagtgctgacccagactccatcccctgtgtctgcagctgtgggaggcacagtcaccatcaactgccagggcagtcagagtctttataataacaacgccttttcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatgatgcttccactctggcgtctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagtggcgtgcagtgtgcagatgctgccacttactactgtcaaggcgaatttagttgtggtgatgttgattgtattgctttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:632)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:630的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:640)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO:614, SEQ ID NO:616 and SEQ ID NO:618 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 601 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 602; and/or SEQ ID NO: 634, One or more of the polynucleotide sequences of SEQ ID NO: 636 and SEQ ID NO: 638, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 621 or the variable light chain sequence of SEQ ID NO: 622 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO:613, SEQ ID NO:615, SEQ ID NO:617 and SEQ ID NO:619 One or more in the acid sequence, it corresponds to the heavy chain sequence of encoding SEQIDNO:601 or the polynucleotide of framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:602; And/or SEQIDNO: 633, one or more of the polynucleotide sequences of SEQIDNO:635, SEQIDNO:637 and SEQIDNO:639, which correspond to the light chain sequence of SEQIDNO:621 or the framework region of the variable light chain sequence of SEQIDNO:622 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:611 encoding the heavy chain sequence of SEQ ID NO:601; the polynucleotide SEQ ID NO:612 encoding the variable heavy chain sequence of SEQ ID NO:602; the polynucleotide encoding the light chain sequence of SEQ ID NO:621 Nucleotide SEQIDNO:631; Polynucleotide SEQIDNO:632 encoding the variable light chain sequence of SEQIDNO:622; The complementarity determining region (SEQIDNO:614 of the heavy chain sequence of encoding SEQIDNO:601 or the variable heavy chain sequence of SEQIDNO:602 , SEQIDNO:616 and SEQIDNO:618) polynucleotides; encoding the light chain sequence of SEQIDNO:621 or the complementarity determining region (SEQIDNO:634, SEQIDNO:636 and SEQIDNO:638) of the variable light chain sequence of SEQIDNO:622 A polynucleotide; a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 601 or the framework region (SEQ ID NO: 613, SEQ ID NO: 615, SEQ ID NO: 617 and SEQ ID NO: 619) of the variable heavy chain sequence of SEQ ID NO: 602; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 621 or the framework region (SEQ ID NO: 633, SEQ ID NO: 635, SEQ ID NO: 637 and SEQ ID NO: 639) of the variable light chain sequence of SEQ ID NO: 622.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab16, the polynucleotide encoding the full-length Ab16 antibody comprises or consists of the polynucleotide SEQ ID NO:611 encoding the heavy chain sequence of SEQ ID NO:601 and the polynucleotide SEQ ID NO:621 encoding the light chain sequence: 631.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl6 after expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, anti-HGF antibodies such as Ab16 or its Fab fragments can be expressed in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab17

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:641的重链序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcccgcctgggacacccctgacactcacctgcacagtctctggaatcgacctcagtagctatgcaatgggctgggtccgccaggctccagggaaggggctggaatacatcggaatgattgatgttagtggtagcacgtactacgcggactgggcgaaaggccgactcaccatctccaaaaccccgaccacggtggatctggaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagattgattgacgagcgttcaacttatagttatgcttttgacttgtggggccaaggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggact ggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:651)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:642的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcccgcctgggacacccctgacactcacctgcacagtctctggaatcgacctcagtagctatgcaatgggctgggtccgccaggctccagggaaggggctggaatacatcggaatgattgatgttagtggtagcacgtactacgcggactgggcgaaaggccgactcaccatctccaaaaccccgaccacggtggatctggaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagattgattgacgagcgttcaacttatagttatgcttttgacttgtggggccaaggcaccctcgtcaccgtctcgagc(SEQIDNO:652)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:650的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 660).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:661的轻链多肽序列的以下多核苷酸序列或者由其组成：caagtgctgacccagactccatcccctgtgtctgcagctgtgggaggcacagtcaccatcaactgccaggccagtcagagtttttataataacggcgccttttcctggtatcagcagaaaccagggcagcctcccaagctcctgatctacgatgcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcggcgtacagtgtggagatgctgccacttactactgtcaaggcgaatttagttgtggtagtgctgattgtgttgctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:671)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:662的可变轻链多肽序列的以下多核苷酸序列或者由其组成：caagtgctgacccagactccatcccctgtgtctgcagctgtgggaggcacagtcaccatcaactgccaggccagtcagagtttttataataacggcgccttttcctggtatcagcagaaaccagggcagcctcccaagctcctgatctacgatgcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcggcgtacagtgtggagatgctgccacttactactgtcaaggcgaatttagttgtggtagtgctgattgtgttgctttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:672)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:670的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:680)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO:654, SEQ ID NO:656 and SEQ ID NO:658 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 641 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 642; and/or SEQ ID NO: 674, One or more of the polynucleotide sequences of SEQ ID NO:676 and SEQ ID NO:678, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO:661 or the variable light chain sequence of SEQ ID NO:662 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 653, SEQ ID NO: 655, SEQ ID NO: 657 and SEQ ID NO: 659 One or more of the acid sequences corresponding to the polynucleotide encoding the heavy chain sequence of SEQ ID NO: 641 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 642; and/or SEQ ID NO: 673, one or more of the polynucleotide sequences of SEQIDNO:675, SEQIDNO:677 and SEQIDNO:679, which correspond to the light chain sequence of SEQIDNO:661 or the framework region of the variable light chain sequence of SEQIDNO:662 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:651 encoding the heavy chain sequence of SEQ ID NO:641; the polynucleotide SEQ ID NO:652 encoding the variable heavy chain sequence of SEQ ID NO:642; the polynucleotide encoding the light chain sequence of SEQ ID NO:661 Nucleotide SEQIDNO:671; Polynucleotide SEQIDNO:672 encoding the variable light chain sequence of SEQIDNO:662; The complementarity determining region (SEQIDNO:654 of the heavy chain sequence of encoding SEQIDNO:641 or the variable heavy chain sequence of SEQIDNO:642 , SEQIDNO:656 and SEQIDNO:658) polynucleotides; encoding the light chain sequence of SEQIDNO:661 or the complementarity determining region (SEQIDNO:674, SEQIDNO:676 and SEQIDNO:678) of the variable light chain sequence of SEQIDNO:662 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 641 or the framework region (SEQ ID NO: 653, SEQ ID NO: 655, SEQ ID NO: 657 and SEQ ID NO: 659) of the variable heavy chain sequence of SEQ ID NO: 642; and encoding SEQ ID NO A polynucleotide of the light chain sequence of 661 or the framework region (SEQ ID NO: 673, SEQ ID NO: 675, SEQ ID NO: 677 and SEQ ID NO: 679) of the variable light chain sequence of SEQ ID NO: 662.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Abl7, the polynucleotide encoding the full-length Ab17 antibody comprises or consists of the polynucleotide SEQ ID NO:651 encoding the heavy chain sequence of SEQ ID NO:641 and the polynucleotide SEQ ID NO:661 encoding the light chain sequence: 671.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl7 after expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, anti-HGF antibodies such as Ab17 or its Fab fragments can be expressed via Ab17 polynucleotides in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab18

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:681的重链序列的以下多核苷酸序列或者由其组成：cagtcgctggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagcctctggattctccctcagcagctacgacatgagctgggtccgccaggctccagggaaggggctggaatggatcggaatcatttatgctggtagtgctagcacatggttcgcgagctgggtgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatgaccagtctgacaaccgaggacacggccacctatttctgtgccagagtgggttatagtggttatggttatgatgataatttggacatgtggggccaaggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccagg actggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:691)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:682的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcgctggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagcctctggattctccctcagcagctacgacatgagctgggtccgccaggctccagggaaggggctggaatggatcggaatcatttatgctggtagtgctagcacatggttcgcgagctgggtgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatgaccagtctgacaaccgaggacacggccacctatttctgtgccagagtgggttatagtggttatggttatgatgataatttggacatgtggggccaaggcaccctcgtcaccgtctcgagc(SEQIDNO:692)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:690的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 700).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:701的轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagcactgcattagcctggtatcagcagaaaccagggcagcgtcccaagctcctgatctacgatgcatcgaaactggcatctggggtctcatcgcggttcaaaggcagtggatctggggcacagttcactctcaccatcagcggcgtggagtgtgccgatgctgccacttactactgtcaccagggttatagtagtagtaatgttgataatactttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:711)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:702的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagcactgcattagcctggtatcagcagaaaccagggcagcgtcccaagctcctgatctacgatgcatcgaaactggcatctggggtctcatcgcggttcaaaggcagtggatctggggcacagttcactctcaccatcagcggcgtggagtgtgccgatgctgccacttactactgtcaccagggttatagtagtagtaatgttgataatactttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:712)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:710的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:720)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO:694, SEQ ID NO:696 and SEQ ID NO:698 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 681 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 682; and/or SEQ ID NO: 714, One or more of the polynucleotide sequences of SEQ ID NO: 716 and SEQ ID NO: 718, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 701 or the variable light chain sequence of SEQ ID NO: 702 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO:693, SEQ ID NO:695, SEQ ID NO:697 and SEQ ID NO:699 One or more of acid sequences corresponding to the polynucleotide encoding the heavy chain sequence of SEQ ID NO: 681 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 682; and/or SEQ ID NO: 713, one or more of the polynucleotide sequences of SEQIDNO:715, SEQIDNO:717 and SEQIDNO:719, which correspond to the light chain sequence of SEQIDNO:701 or the framework region of the variable light chain sequence of SEQIDNO:702 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:691 encoding the heavy chain sequence of SEQ ID NO:681; the polynucleotide SEQ ID NO:692 encoding the variable heavy chain sequence of SEQ ID NO:682; the polynucleotide encoding the light chain sequence of SEQ ID NO:701 Nucleotide SEQIDNO:711; polynucleotide SEQIDNO:712 encoding the variable light chain sequence of SEQIDNO:702; the complementarity determining region (SEQIDNO:694) of the heavy chain sequence encoding SEQIDNO:681 or the variable heavy chain sequence of SEQIDNO:682 , SEQIDNO:696 and SEQIDNO:698) polynucleotides; encoding the light chain sequence of SEQIDNO:701 or the complementary determining region (SEQIDNO:714, SEQIDNO:716 and SEQIDNO:718) of the variable light chain sequence of SEQIDNO:702 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 681 or the framework region (SEQ ID NO: 693, SEQ ID NO: 695, SEQ ID NO: 697 and SEQ ID NO: 699) of the variable heavy chain sequence of SEQ ID NO: 682; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 701 or the framework region (SEQ ID NO: 713, SEQ ID NO: 715, SEQ ID NO: 717 and SEQ ID NO: 719) of the variable light chain sequence of SEQ ID NO: 702.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab18, the polynucleotide encoding the full-length Ab18 antibody comprises or consists of the polynucleotide SEQ ID NO:691 encoding the heavy chain sequence of SEQ ID NO:681 and the polynucleotide SEQ ID NO:691 encoding the light chain sequence of SEQ ID NO:701: 711.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl8 after expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, anti-HGF antibodies such as Ab18 or its Fab fragments can be expressed via Ab18 polynucleotides in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab19

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:721的重链序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagcctctggattctccctcagtaattattggatgggctgggtccgccaggctccaggggaggggctggaatggatcggaaccattagttatgatggtaacacatactacgcgagctgggcaaaaggccgcttcaccatctcccgaacctcgaccacggtggatctgaaaatgaccagtctgacgaccgaggacacggccatctatttctgtgccacagtcaattatcctgattatagtactggtgcctttaacatctggggcccaggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggc tgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:731)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:722的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagcctctggattctccctcagtaattattggatgggctgggtccgccaggctccaggggaggggctggaatggatcggaaccattagttatgatggtaacacatactacgcgagctgggcaaaaggccgcttcaccatctcccgaacctcgaccacggtggatctgaaaatgaccagtctgacgaccgaggacacggccatctatttctgtgccacagtcaattatcctgattatagtactggtgcctttaacatctggggcccaggcaccctcgtcaccgtctcgagc(SEQIDNO:732)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:730的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 740).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:741的轻链多肽序列的以下多核苷酸序列或者由其组成：gatgttgtgatgacccagactccagcctccgtgtctgaacctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattgataactacttagcctggtatcagcagaaaccagggcagcgtcccaggctcctgatctattatacatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacagagtacactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaatttactgcttattatagtacttatattggagctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:751)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:742的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gatgttgtgatgacccagactccagcctccgtgtctgaacctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattgataactacttagcctggtatcagcagaaaccagggcagcgtcccaggctcctgatctattatacatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacagagtacactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaatttactgcttattatagtacttatattggagctttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:752)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:750的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:760)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 734, SEQ ID NO: 736 and SEQ ID NO: 738 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 721 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 722; and/or SEQ ID NO: 754, One or more of the polynucleotide sequences of SEQ ID NO: 756 and SEQ ID NO: 758, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 741 or the variable light chain sequence of SEQ ID NO: 742 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 733, SEQ ID NO: 735, SEQ ID NO: 737 and SEQ ID NO: 739 One or more in the acid sequence, it corresponds to the polynucleotide of the heavy chain sequence of encoding SEQIDNO:721 or the frame region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:722; And/or SEQIDNO: 753, one or more of the polynucleotide sequences of SEQIDNO:755, SEQIDNO:757 and SEQIDNO:759, which correspond to the light chain sequence of SEQIDNO:741 or the framework region of the variable light chain sequence of SEQIDNO:742 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:731 encoding the heavy chain sequence of SEQ ID NO:721; the polynucleotide SEQ ID NO:732 encoding the variable heavy chain sequence of SEQ ID NO:722; the polynucleotide encoding the light chain sequence of SEQ ID NO:741 Nucleotide SEQIDNO:751; Polynucleotide SEQIDNO:752 encoding the variable light chain sequence of SEQIDNO:742; The complementarity determining region (SEQIDNO:734 of the heavy chain sequence of encoding SEQIDNO:721 or the variable heavy chain sequence of SEQIDNO:722 , SEQIDNO:736 and SEQIDNO:738) polynucleotides; encoding the light chain sequence of SEQIDNO:741 or the complementarity determining region (SEQIDNO:754, SEQIDNO:756 and SEQIDNO:758) of the variable light chain sequence of SEQIDNO:742 A polynucleotide; a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 721 or the framework region (SEQ ID NO: 733, SEQ ID NO: 735, SEQ ID NO: 737 and SEQ ID NO: 739) of the variable heavy chain sequence of SEQ ID NO: 722; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 741 or the framework region (SEQ ID NO: 753, SEQ ID NO: 755, SEQ ID NO: 757 and SEQ ID NO: 759) of the variable light chain sequence of SEQ ID NO: 742.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab19, the polynucleotide encoding the full-length Ab19 antibody comprises or consists of the polynucleotide SEQ ID NO:731 encoding the heavy chain sequence of SEQ ID NO:721 and the polynucleotide SEQ ID NO:741 encoding the light chain sequence: 751.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Abl9 after expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, anti-HGF antibodies such as Ab19 or its Fab fragments can be expressed via Ab19 polynucleotides in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab20

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:761的重链序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtaactattggatgggctgggtccgtcaggctccagggaaggggctggagtggatcggaaccattagttatgatggtaacacatactacgcgagcagcgcaaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctacagtcaattatcctgattatagtactggtgcctttaacatctggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcacc aggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:771)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:762的可变重链多肽序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtaactattggatgggctgggtccgtcaggctccagggaaggggctggagtggatcggaaccattagttatgatggtaacacatactacgcgagcagcgcaaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctacagtcaattatcctgattatagtactggtgcctttaacatctggggccaagggaccctcgtcaccgtctcgagc(SEQIDNO:772)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:770的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 780).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:781的轻链多肽序列的以下多核苷酸序列或者由其组成：gacatccagatgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcacttgccaggccagtcagagcattgataactacttagcctggtatcagcagaaaccagggaaagttcctaagctcctgatctattatacatccactctggcatctggggtcccatctcgtttcagtggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctgaagatgttgcaacttattactgtcaattcactgcttattatagtacttacattggagctttcggcggaggaaccaaggtggaaatcaaacgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:791)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:782的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gacatccagatgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcacttgccaggccagtcagagcattgataactacttagcctggtatcagcagaaaccagggaaagttcctaagctcctgatctattatacatccactctggcatctggggtcccatctcgtttcagtggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctgaagatgttgcaacttattactgtcaattcactgcttattatagtacttacattggagctttcggcggaggaaccaaggtggaaatcaaacgt(SEQIDNO:792)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:790的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:800)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO:774, SEQ ID NO:776 and SEQ ID NO:778 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 761 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 762; and/or SEQ ID NO: 794, One or more of the polynucleotide sequences of SEQ ID NO: 796 and SEQ ID NO: 798, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 781 or the variable light chain sequence of SEQ ID NO: 782 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO:773, SEQ ID NO:775, SEQ ID NO:777 and SEQ ID NO:779 One or more in the acid sequence, it corresponds to the polynucleotide of the heavy chain sequence of encoding SEQIDNO:761 or the frame region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:762; And/or SEQIDNO: 793, one or more of the polynucleotide sequences of SEQIDNO:795, SEQIDNO:797 and SEQIDNO:799, which correspond to the light chain sequence of SEQIDNO:781 or the framework region of the variable light chain sequence of SEQIDNO:782 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:771 encoding the heavy chain sequence of SEQ ID NO:761; the polynucleotide SEQ ID NO:772 encoding the variable heavy chain sequence of SEQ ID NO:762; the polynucleotide encoding the light chain sequence of SEQ ID NO:781 Nucleotide SEQIDNO:791; Polynucleotide SEQIDNO:792 encoding the variable light chain sequence of SEQIDNO:782; The complementarity determining region (SEQIDNO:774 of the heavy chain sequence of encoding SEQIDNO:761 or the variable heavy chain sequence of SEQIDNO:762 , SEQIDNO:776 and SEQIDNO:778) polynucleotides; encoding the light chain sequence of SEQIDNO:781 or the complementarity determining region (SEQIDNO:794, SEQIDNO:796 and SEQIDNO:798) of the variable light chain sequence of SEQIDNO:782 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 761 or the framework region (SEQ ID NO: 773, SEQ ID NO: 775, SEQ ID NO: 777 and SEQ ID NO: 779) of the variable heavy chain sequence of SEQ ID NO: 762; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 781 or the framework region (SEQ ID NO: 793, SEQ ID NO: 795, SEQ ID NO: 797 and SEQ ID NO: 799) of the variable light chain sequence of SEQ ID NO: 782.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Ab20, the polynucleotide encoding the full-length Ab20 antibody comprises or consists of the polynucleotide SEQ ID NO: 771 encoding the heavy chain sequence of SEQ ID NO: 761 and the polynucleotide SEQ ID NO: 781 encoding the light chain sequence: 791.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab20 following expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, an anti-HGF antibody such as Ab20 or its Fab fragment can be expressed via Ab20 polynucleotide in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab21

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:801的重链序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcaccgtctctggattctccctcagtacctactacatgagctgggtccgccaggctccagggaaggggctggaatggatcggaatcatttatgttagtggtatcacggactacgcgaggtgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatgaccagtctgacaaccgaggacacggccacctatttctgtgccagacatattgatagtagtggctgggatggactgggcatctggggccaaggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctga atggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:811)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:802的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcaccgtctctggattctccctcagtacctactacatgagctgggtccgccaggctccagggaaggggctggaatggatcggaatcatttatgttagtggtatcacggactacgcgaggtgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatgaccagtctgacaaccgaggacacggccacctatttctgtgccagacatattgatagtagtggctgggatggactgggcatctggggccaaggcaccctcgtcaccgtctcgagc(SEQIDNO:812)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:810的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 820).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:821的轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtgagagcattagtagctacttaaactggtatcagcagaaactagggcagcctcccaagctcctgatctacagggcatccactctgacatctggggtctcatcaaggttcaaaggcagtggatctgggacagagtacactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcagcagacttatggttatagtgatactgataattctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:831)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:822的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtgagagcattagtagctacttaaactggtatcagcagaaactagggcagcctcccaagctcctgatctacagggcatccactctgacatctggggtctcatcaaggttcaaaggcagtggatctgggacagagtacactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcagcagacttatggttatagtgatactgataattctttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:832)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:830的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:840)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO:814, SEQ ID NO:816 and SEQ ID NO:818 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 801 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 802; and/or SEQ ID NO: 834, One or more of the polynucleotide sequences of SEQ ID NO: 836 and SEQ ID NO: 838, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 821 or the variable light chain sequence of SEQ ID NO: 822 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO:813, SEQ ID NO:815, SEQ ID NO:817 and SEQ ID NO:819 One or more in the acid sequence, it corresponds to the heavy chain sequence of encoding SEQIDNO:801 or the polynucleotide of framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:802; And/or SEQIDNO: 833, one or more of the polynucleotide sequences of SEQIDNO:835, SEQIDNO:837 and SEQIDNO:839, which correspond to the light chain sequence of SEQIDNO:821 or the framework region of the variable light chain sequence of SEQIDNO:822 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:811 encoding the heavy chain sequence of SEQ ID NO:801; the polynucleotide SEQ ID NO:812 encoding the variable heavy chain sequence of SEQ ID NO:802; the polynucleotide encoding the light chain sequence of SEQ ID NO:821 Nucleotide SEQIDNO:831; Polynucleotide SEQIDNO:832 encoding the variable light chain sequence of SEQIDNO:822; The complementarity determining region (SEQIDNO:814 of the heavy chain sequence of encoding SEQIDNO:801 or the variable heavy chain sequence of SEQIDNO:802 , SEQIDNO:816 and SEQIDNO:818) polynucleotides; encoding the light chain sequence of SEQIDNO:821 or the complementarity determining region (SEQIDNO:834, SEQIDNO:836 and SEQIDNO:838) of the variable light chain sequence of SEQIDNO:822 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 801 or the framework region (SEQ ID NO: 813, SEQ ID NO: 815, SEQ ID NO: 817 and SEQ ID NO: 819) of the variable heavy chain sequence of SEQ ID NO: 802; and encoding SEQ ID NO A polynucleotide of the light chain sequence of SEQ ID NO:821 or the framework region (SEQ ID NO:833, SEQ ID NO:835, SEQ ID NO:837 and SEQ ID NO:839) of the variable light chain sequence of SEQ ID NO:822.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab21, the polynucleotide encoding the full-length Ab21 antibody comprises or consists of the polynucleotide SEQ ID NO:811 encoding the heavy chain sequence of SEQ ID NO:801 and the polynucleotide SEQ ID NO:821 encoding the light chain sequence: 831.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab21 following expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, an anti-HGF antibody such as Ab21 or its Fab fragment can be expressed via Ab21 polynucleotide in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab23

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:841的重链序列的以下多核苷酸序列或者由其组成：cagtcgctggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagcctctggattcactatcggtcgctactacatgagctgggtccgccaggctccagggaaggggctggaatggatcggaatcatttatactcatggtgttaacccagactacgcgagctgggcgaaaggccgattcaccatctccagaccctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagagtgggtggttttaatgactactctgacatttggggcccaggcaccctggtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatg gcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:851)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:842的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcgctggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagcctctggattcactatcggtcgctactacatgagctgggtccgccaggctccagggaaggggctggaatggatcggaatcatttatactcatggtgttaacccagactacgcgagctgggcgaaaggccgattcaccatctccagaccctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagagtgggtggttttaatgactactctgacatttggggcccaggcaccctggtcaccgtctcgagc(SEQIDNO:852)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:850的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 860).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:861的轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtgagagcattagtacctacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctacagggcatccactctggcatctggggtctcatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcggcgtggagtgtgccgatgctgccacttactactgtcaacagggttatagttatagtaatgttgataatgctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:871)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:862的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtgagagcattagtacctacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctacagggcatccactctggcatctggggtctcatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcggcgtggagtgtgccgatgctgccacttactactgtcaacagggttatagttatagtaatgttgataatgctttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:872)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:870的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:880)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO:854, SEQ ID NO:856 and SEQ ID NO:858 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 841 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 842; and/or SEQ ID NO: 874, One or more of the polynucleotide sequences of SEQ ID NO: 876 and SEQ ID NO: 878, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 861 or the variable light chain sequence of SEQ ID NO: 862 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO:853, SEQ ID NO:855, SEQ ID NO:857 and SEQ ID NO:859 One or more in the acid sequence, it corresponds to the heavy chain sequence of encoding SEQIDNO:841 or the polynucleotide of framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:842; And/or SEQIDNO: 873, one or more of the polynucleotide sequences of SEQIDNO:875, SEQIDNO:877 and SEQIDNO:879, which correspond to the light chain sequence of SEQIDNO:861 or the framework region of the variable light chain sequence of SEQIDNO:862 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:851 encoding the heavy chain sequence of SEQ ID NO:841; the polynucleotide SEQ ID NO:852 encoding the variable heavy chain sequence of SEQ ID NO:842; the polynucleotide encoding the light chain sequence of SEQ ID NO:861 Nucleotide SEQIDNO:871; Polynucleotide SEQIDNO:872 encoding the variable light chain sequence of SEQIDNO:862; The complementarity determining region (SEQIDNO:854 of the heavy chain sequence of encoding SEQIDNO:841 or the variable heavy chain sequence of SEQIDNO:842 , SEQIDNO:856 and SEQIDNO:858) polynucleotides; encoding the light chain sequence of SEQIDNO:861 or the complementarity determining region (SEQIDNO:874, SEQIDNO:876 and SEQIDNO:878) of the variable light chain sequence of SEQIDNO:862 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 841 or the framework regions (SEQ ID NO: 853, SEQ ID NO: 855, SEQ ID NO: 857 and SEQ ID NO: 859) of the variable heavy chain sequence of SEQ ID NO: 842; and encoding SEQ ID NO A polynucleotide of the light chain sequence of SEQ ID NO:861 or the framework region (SEQ ID NO:873, SEQ ID NO:875, SEQ ID NO:877 and SEQ ID NO:879) of the variable light chain sequence of SEQ ID NO:862.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab23, the polynucleotide encoding the full-length Ab23 antibody comprises or consists of the polynucleotide SEQ ID NO:851 encoding the heavy chain sequence of SEQ ID NO:841 and the polynucleotide SEQ ID NO:861 encoding the light chain sequence: 871.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab23 after expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, an anti-HGF antibody such as Ab23 or its Fab fragment can be expressed via Ab23 polynucleotide in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab24

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:881的重链序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcggtcgctactacatgagctgggtccgtcaggctccagggaaggggctggagtggatcggaatcatctatactcatggtgttaacccagactacgcgagcagcgcgaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagagtgggtggtttcaatgactactctgacatttggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggact ggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:891)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:882的可变重链多肽序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcggtcgctactacatgagctgggtccgtcaggctccagggaaggggctggagtggatcggaatcatctatactcatggtgttaacccagactacgcgagcagcgcgaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagagtgggtggtttcaatgactactctgacatttggggccaagggaccctcgtcaccgtctcgagc(SEQIDNO:892)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:890的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 900).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:901的轻链多肽序列的以下多核苷酸序列或者由其组成：gacatccagatgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcacttgccaggccagtgagagcattagtacctacttagcctggtatcagcagaaaccagggaaagttcctaagctcctgatctatagggcatccactctggcatctggggtcccatctcgtttcagtggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctgaagatgttgcaacttattactgtcaacagggttatagttatagtaatgttgataatgctttcggcggaggaaccaaggtggaaatcaaacgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:911)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:902的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gacatccagatgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcacttgccaggccagtgagagcattagtacctacttagcctggtatcagcagaaaccagggaaagttcctaagctcctgatctatagggcatccactctggcatctggggtcccatctcgtttcagtggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctgaagatgttgcaacttattactgtcaacagggttatagttatagtaatgttgataatgctttcggcggaggaaccaaggtggaaatcaaacgt(SEQIDNO:912)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:910的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:920)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO:894, SEQ ID NO:896 and SEQ ID NO:898 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 881 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 882; and/or SEQ ID NO: 914, One or more of the polynucleotide sequences of SEQ ID NO: 916 and SEQ ID NO: 918, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 901 or the variable light chain sequence of SEQ ID NO: 902 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO:893, SEQ ID NO:895, SEQ ID NO:897 and SEQ ID NO:899 One or more of the acid sequences corresponding to the polynucleotide encoding the heavy chain sequence of SEQ ID NO: 881 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 882; and/or SEQ ID NO: 913, one or more of the polynucleotide sequences of SEQIDNO:915, SEQIDNO:917 and SEQIDNO:919, which correspond to the light chain sequence of SEQIDNO:901 or the framework region of the variable light chain sequence of SEQIDNO:902 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:891 encoding the heavy chain sequence of SEQ ID NO:881; the polynucleotide SEQ ID NO:892 encoding the variable heavy chain sequence of SEQ ID NO:882; the polynucleotide encoding the light chain sequence of SEQ ID NO:901 Nucleotide of SEQIDNO:911; Polynucleotide SEQIDNO:912 encoding the variable light chain sequence of SEQIDNO:902; The complementarity determining region (SEQIDNO:894 of the heavy chain sequence of encoding SEQIDNO:881 or the variable heavy chain sequence of SEQIDNO:882 , SEQIDNO:896 and SEQIDNO:898) polynucleotides; encoding the light chain sequence of SEQIDNO:901 or the complementary determining region (SEQIDNO:914, SEQIDNO:916 and SEQIDNO:918) of the variable light chain sequence of SEQIDNO:902 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 881 or the framework region (SEQ ID NO: 893, SEQ ID NO: 895, SEQ ID NO: 897 and SEQ ID NO: 899) of the variable heavy chain sequence of SEQ ID NO: 882; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 901 or the framework region (SEQ ID NO: 913, SEQ ID NO: 915, SEQ ID NO: 917 and SEQ ID NO: 919) of the variable light chain sequence of SEQ ID NO: 902.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Ab24, the polynucleotide encoding the full-length Ab24 antibody comprises or consists of the polynucleotide SEQ ID NO:891 encoding the heavy chain sequence of SEQ ID NO:881 and the polynucleotide SEQ ID NO:891 encoding the light chain sequence of SEQ ID NO:901: 911.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab24 after expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, an anti-HGF antibody such as Ab24 or its Fab fragment can be expressed via Ab24 polynucleotide in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab25

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:921的重链序列的以下多核苷酸序列或者由其组成：cagtcgctggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagcctctggattctccctcagtagctatgcaatgggctggttccgccaggctccagggaaggggctggagtggatcgcatacatttttgctagtggtagcacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggagctgaaaatcaccagtctgacaaccgaggacacggccacctatttctgtgccagaggtagtggtgctcgttttttccccaactactttgccatctggggcccaggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggc tgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:931)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:922的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcgctggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagcctctggattctccctcagtagctatgcaatgggctggttccgccaggctccagggaaggggctggagtggatcgcatacatttttgctagtggtagcacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggagctgaaaatcaccagtctgacaaccgaggacacggccacctatttctgtgccagaggtagtggtgctcgttttttccccaactactttgccatctggggcccaggcaccctcgtcaccgtctcgagc(SEQIDNO:932)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:930的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 940).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:941的轻链多肽序列的以下多核苷酸序列或者由其组成：caagtgctgacccagactgcatcgtccgtgtctgcagctgtgggaggcacagtcaccatcagttgccagtccagtcagagtgttactaataacaacgacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctaccaggcatccaaactggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcgacctggagtgtgacgatgctgccacttactactgtcaaggcagttatagtggtggtatttgtgctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:951)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:942的可变轻链多肽序列的以下多核苷酸序列或者由其组成：caagtgctgacccagactgcatcgtccgtgtctgcagctgtgggaggcacagtcaccatcagttgccagtccagtcagagtgttactaataacaacgacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctaccaggcatccaaactggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcgacctggagtgtgacgatgctgccacttactactgtcaaggcagttatagtggtggtatttgtgctttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:952)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:950的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:960)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 934, SEQ ID NO: 936 and SEQ ID NO: 938 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 921 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 922; and/or SEQ ID NO: 954, One or more of the polynucleotide sequences of SEQ ID NO: 956 and SEQ ID NO: 958, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 941 or the variable light chain sequence of SEQ ID NO: 942 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 933, SEQ ID NO: 935, SEQ ID NO: 937 and SEQ ID NO: 939 One or more of acid sequences corresponding to the polynucleotide encoding the heavy chain sequence of SEQ ID NO: 921 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 922; and/or SEQ ID NO: 953, one or more of the polynucleotide sequences of SEQIDNO:955, SEQIDNO:957 and SEQIDNO:959, which correspond to the light chain sequence of SEQIDNO:941 or the framework region of the variable light chain sequence of SEQIDNO:942 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:931 encoding the heavy chain sequence of SEQ ID NO:921; the polynucleotide SEQ ID NO:932 encoding the variable heavy chain sequence of SEQ ID NO:922; the polynucleotide encoding the light chain sequence of SEQ ID NO:941 Nucleotide SEQIDNO:951; polynucleotide SEQIDNO:952 encoding the variable light chain sequence of SEQIDNO:942; the complementarity determining region (SEQIDNO:934) of the heavy chain sequence encoding SEQIDNO:921 or the variable heavy chain sequence of SEQIDNO:922 , SEQIDNO:936 and SEQIDNO:938) polynucleotides; encoding the light chain sequence of SEQIDNO:941 or the complementarity determining region (SEQIDNO:954, SEQIDNO:956 and SEQIDNO:958) of the variable light chain sequence of SEQIDNO:942 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 921 or the framework region (SEQ ID NO: 933, SEQ ID NO: 935, SEQ ID NO: 937 and SEQ ID NO: 939) of the variable heavy chain sequence of SEQ ID NO: 922; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 941 or the framework region (SEQ ID NO: 953, SEQ ID NO: 955, SEQ ID NO: 957 and SEQ ID NO: 959) of the variable light chain sequence of SEQ ID NO: 942.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab25, the polynucleotide encoding the full-length Ab25 antibody comprises or consists of the polynucleotide SEQ ID NO:931 encoding the heavy chain sequence of SEQ ID NO:921 and the polynucleotide SEQ ID NO:941 encoding the light chain sequence: 951.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab25 following expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, an anti-HGF antibody such as Ab25 or its Fab fragment can be expressed via Ab25 polynucleotide in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab26

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:961的重链序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtacctatacaatgaactgggtccgccaggctccagggaaggggctggaatacatcggattcattagtagtagtagtagcatagattatgtgagttgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagagatttttatgctgattatattggtggtggttatccttacatctggggcccgggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggact ggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:971)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:962的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtacctatacaatgaactgggtccgccaggctccagggaaggggctggaatacatcggattcattagtagtagtagtagcatagattatgtgagttgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagagatttttatgctgattatattggtggtggttatccttacatctggggcccgggcaccctcgtcaccgtctcgagc(SEQIDNO:972)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:970的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgag gctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 980).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:981的轻链多肽序列的以下多核苷酸序列或者由其组成：gccgatgttgtgatgacccagactccagcctccgtgtctgaacctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtagctacttatcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatggtgcatccaaactgacatctggggtcccatcgcggttcaaaggcagtggatctgggacagagtacactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaagcaattatgatatttatagttatgctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:991)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:982的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gccgatgttgtgatgacccagactccagcctccgtgtctgaacctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtagctacttatcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatggtgcatccaaactgacatctggggtcccatcgcggttcaaaggcagtggatctgggacagagtacactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaagcaattatgatatttatagttatgctttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:992)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:990的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:1000)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 974, SEQ ID NO: 976 and SEQ ID NO: 978 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 961 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 962; and/or SEQ ID NO: 994, One or more of the polynucleotide sequences of SEQ ID NO: 996 and SEQ ID NO: 998, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 981 or the variable light chain sequence of SEQ ID NO: 982 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 973, SEQ ID NO: 975, SEQ ID NO: 977 and SEQ ID NO: 979 One or more of acid sequences corresponding to the polynucleotide encoding the heavy chain sequence of SEQ ID NO: 961 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 962; and/or SEQ ID NO: 993, one or more of the polynucleotide sequences of SEQIDNO:995, SEQIDNO:997 and SEQIDNO:999, which correspond to the light chain sequence of SEQIDNO:981 or the framework region of the variable light chain sequence of SEQIDNO:982 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO:971 encoding the heavy chain sequence of SEQ ID NO:961; the polynucleotide SEQ ID NO:972 encoding the variable heavy chain sequence of SEQ ID NO:962; the polynucleotide encoding the light chain sequence of SEQ ID NO:981 Nucleotide of SEQIDNO:991; polynucleotide SEQIDNO:992 encoding the variable light chain sequence of SEQIDNO:982; the complementarity determining region (SEQIDNO:974) of the heavy chain sequence encoding SEQIDNO:961 or the variable heavy chain sequence of SEQIDNO:962 , SEQIDNO:976 and SEQIDNO:978) polynucleotides; encoding the light chain sequence of SEQIDNO:981 or the complementarity determining region (SEQIDNO:994, SEQIDNO:996 and SEQIDNO:998) of the variable light chain sequence of SEQIDNO:982 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 961 or the framework region (SEQ ID NO: 973, SEQ ID NO: 975, SEQ ID NO: 977 and SEQ ID NO: 979) of the variable heavy chain sequence of SEQ ID NO: 962; and encoding SEQ ID NO A polynucleotide of the light chain sequence of 981 or the framework region (SEQ ID NO: 993, SEQ ID NO: 995, SEQ ID NO: 997 and SEQ ID NO: 999) of the variable light chain sequence of SEQ ID NO: 982.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab26, the polynucleotide encoding the full-length Ab26 antibody comprises or consists of the polynucleotide SEQ ID NO:971 encoding the heavy chain sequence of SEQ ID NO:961 and the polynucleotide SEQ ID NO:981 encoding the light chain sequence: 991.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab26 following expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, an anti-HGF antibody such as Ab26 or its Fab fragment can be expressed via Ab26 polynucleotide in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab27

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:1001的重链序列的以下多核苷酸序列或者由其组成：cagtcgttggaggagtccgggggagacctggtcaagcctggaggaaccctgacactcacctgcacagcctctggattctccttcagtgacgaccactacatgtgctgggtccgccaggctccagggaaggggctgcagtggatcgcatgcatgtatgttggtagtagtggtgccacttattacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgtcgaccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacctatttctgtgcgagagatgattggactagttattatgcgtgggggtattgggccttgtggggcccgggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtc ctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:1011)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:1002的可变重链多肽序列的以下多核苷酸序列或者由其组成：cagtcgttggaggagtccgggggagacctggtcaagcctggaggaaccctgacactcacctgcacagcctctggattctccttcagtgacgaccactacatgtgctgggtccgccaggctccagggaaggggctgcagtggatcgcatgcatgtatgttggtagtagtggtgccacttattacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgtcgaccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacctatttctgtgcgagagatgattggactagttattatgcgtgggggtattgggccttgtggggcccgggcaccctcgtcaccgtctcgagc(SEQIDNO:1012)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:1010的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatga ggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 1020).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:1021的轻链多肽序列的以下多核苷酸序列或者由其组成：gctgacattgtgatgacccagaatccagcctccgtgtctgaacctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcgttaatagttggttatcctggtatcagcagaaaccagggcagcctcccaagttcctgatctacaaggcatccactctggcatctggggtctcatcgcggttcaaaggcagtgggattgggacagagttcactctcaccatcagcgacctggagtgtgccgatgctgccacgtactattgccaatttagtaatagtggtactatttatgggagtggtttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:1031)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:1022的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gctgacattgtgatgacccagaatccagcctccgtgtctgaacctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcgttaatagttggttatcctggtatcagcagaaaccagggcagcctcccaagttcctgatctacaaggcatccactctggcatctggggtctcatcgcggttcaaaggcagtgggattgggacagagttcactctcaccatcagcgacctggagtgtgccgatgctgccacgtactattgccaatttagtaatagtggtactatttatgggagtggtttcggcggagggaccgaggtggtggtcaaacgt(SEQIDNO:1032)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:1030的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:1040)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 1014, SEQ ID NO: 1016 and SEQ ID NO: 1018 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 1001 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 1002; and/or SEQ ID NO: 1034, One or more of the polynucleotide sequences of SEQ ID NO: 1036 and SEQ ID NO: 1038, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 1021 or the variable light chain sequence of SEQ ID NO: 1022 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 1013, SEQ ID NO: 1015, SEQ ID NO: 1017 and SEQ ID NO: 1019 One or more of acid sequences corresponding to the polynucleotide encoding the heavy chain sequence of SEQ ID NO: 1001 or the framework region (FR or constant region) of the variable heavy chain sequence of SEQ ID NO: 1002; and/or SEQ ID NO: One or more of the polynucleotide sequences of 1033, SEQIDNO:1035, SEQIDNO:1037 and SEQIDNO:1039, which correspond to the light chain sequence of SEQIDNO:1021 or the framework region of the variable light chain sequence of SEQIDNO:1022 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO: 1011 encoding the heavy chain sequence of SEQ ID NO: 1001; the polynucleotide SEQ ID NO: 1012 encoding the variable heavy chain sequence of SEQ ID NO: 1002; the polynucleotide encoding the light chain sequence of SEQ ID NO: 1021 Nucleotide SEQIDNO:1031; polynucleotide SEQIDNO:1032 encoding the variable light chain sequence of SEQIDNO:1022; the complementarity determining region (SEQIDNO:1014) of the heavy chain sequence encoding SEQIDNO:1001 or the variable heavy chain sequence of SEQIDNO:1002 , SEQIDNO:1016 and SEQIDNO:1018) polynucleotides; encoding the light chain sequence of SEQIDNO:1021 or the complementarity determining region (SEQIDNO:1034, SEQIDNO:1036 and SEQIDNO:1038) of the variable light chain sequence of SEQIDNO:1022 A polynucleotide; a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 1001 or the framework region (SEQ ID NO: 1013, SEQ ID NO: 1015, SEQ ID NO: 1017 and SEQ ID NO: 1019) of the variable heavy chain sequence of SEQ ID NO: 1002; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 1021 or the framework region (SEQ ID NO: 1033, SEQ ID NO: 1035, SEQ ID NO: 1037 and SEQ ID NO: 1039) of the variable light chain sequence of SEQ ID NO: 1022.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. Regarding antibody Ab27, the polynucleotide encoding the full-length Ab27 antibody comprises or consists of the polynucleotide SEQ ID NO: 1011 encoding the heavy chain sequence of SEQ ID NO: 1001 and the polynucleotide SEQ ID NO: 1021 encoding the light chain sequence: 1031.

Another embodiment of the invention encompasses the incorporation of these polynucleotides into expression vectors for use in mammalian cells such as CHO, NSO, HEK-293 or in fungal, insect or microbial systems such as yeast cells, such as Saccharomyces spp. expressed in yeast). Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion (eg papain) of Ab27 following expression of the full-length polynucleotide in a suitable host. In another embodiment of the present invention, an anti-HGF antibody such as Ab27 or its Fab fragment can be expressed via Ab27 polynucleotide in mammalian cells such as CHO, NSO or HEK293 cells, fungi, insects or microbial systems (such as yeast cells ( For example, produced by expression in diploid yeast, such as diploid Pichia and other yeast strains). Suitable Pichia species include, but are not limited to, Pichia pastoris.

Antibody Ab28

In one embodiment, the present invention also relates to polynucleotides encoding antibody polypeptides having binding specificity for HGF.在本发明的一个实施方案中，本发明的多核苷酸包含编码SEQIDNO:1041的重链序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcaatgcaataagctgggtccgtcaggctccagggaaggggctggagtgggtcggagtcatttatgttattggtgtcactgactacgcgagctctgcgcaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagagtttatgattctggctggaatcactttaacttgtggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacgcgagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggac tggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa(SEQIDNO:1051)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:1042的可变重链多肽序列的以下多核苷酸序列或者由其组成：gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcaatgcaataagctgggtccgtcaggctccagggaaggggctggagtgggtcggagtcatttatgttattggtgtcactgactacgcgagctctgcgcaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagagtttatgattctggctggaatcactttaacttgtggggccaagggaccctcgtcaccgtctcgagc(SEQIDNO:1052)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:1050的恒定重链多肽序列的以下多核苷酸序列或者由其组成：gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacgcgagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatga ggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa (SEQ ID NO: 1060).

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:1061的轻链多肽序列的以下多核苷酸序列或者由其组成：gacatccagatgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgccaggccagtcagagcattagcagttggttagcctggtatcagcagaaaccaggaaaagcccctaagctcctgatctatgaagcatccaaactggcatctggagtcccatcaaggttcagcggcagtggatctggaacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttattactgccaacaggcttatagtgttgccaatgttgataatgctttcggcggaggaaccaaggtggaaatcaaacgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:1071)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:1062的可变轻链多肽序列的以下多核苷酸序列或者由其组成：gacatccagatgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgccaggccagtcagagcattagcagttggttagcctggtatcagcagaaaccaggaaaagcccctaagctcctgatctatgaagcatccaaactggcatctggagtcccatcaaggttcagcggcagtggatctggaacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttattactgccaacaggcttatagtgttgccaatgttgataatgctttcggcggaggaaccaaggtggaaatcaaacgt(SEQIDNO:1072)。

在本发明的另一实施方案中，本发明的多核苷酸包含编码SEQIDNO:1070的恒定轻链多肽序列的以下多核苷酸序列或者由其组成：acggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt(SEQIDNO:1080)。

In another embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the following: in the polynucleotide sequences of SEQ ID NO: 1054, SEQ ID NO: 1056 and SEQ ID NO: 1058 One or more that correspond to a polynucleotide encoding the heavy chain sequence of SEQ ID NO: 1041 or the complementarity determining region (CDR or hypervariable region) of the variable heavy chain sequence of SEQ ID NO: 1042; and/or SEQ ID NO: 1074, One or more of the polynucleotide sequences of SEQ ID NO: 1076 and SEQ ID NO: 1078, which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the light chain sequence of SEQ ID NO: 1061 or the variable light chain sequence of SEQ ID NO: 1062 ); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the CDRs set forth above, the variable heavy chain and variable light chain sequences and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

In another embodiment of the invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises or consists of the polynucleotides of SEQ ID NO: 1053, SEQ ID NO: 1055, SEQ ID NO: 1057 and SEQ ID NO: 1059 One or more in the acid sequence, it corresponds to the heavy chain sequence of encoding SEQIDNO:1041 or the polynucleotide of framework region (FR or constant region) of the variable heavy chain sequence of SEQIDNO:1042; And/or SEQIDNO: 1073, one or more of the polynucleotide sequences of SEQ ID NO: 1075, SEQ ID NO: 1077 and SEQ ID NO: 1079, which correspond to the light chain sequence of SEQ ID NO: 1061 or the framework region of the variable light chain sequence of SEQ ID NO: 1062 ( FR or constant region); or a combination of these polynucleotide sequences. In another embodiment of the present invention, the polynucleotide encoding the antibody or fragment thereof of the present invention comprises or consists of the following: encoding the FR, variable heavy chain and variable light chain sequences set forth above and the heavy chain and Combinations of polynucleotides of one or more of the light chain sequences include all such polynucleotides.

The invention also encompasses polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding an antibody fragment with binding specificity for HGF comprises one, two, three or more than three (including all) of the following polynucleotides encoding an antibody fragment ) or consist of: the polynucleotide SEQ ID NO: 1051 encoding the heavy chain sequence of SEQ ID NO: 1041; the polynucleotide SEQ ID NO: 1052 encoding the variable heavy chain sequence of SEQ ID NO: 1042; the polynucleotide encoding the light chain sequence of SEQ ID NO: 1061 Nucleotide SEQIDNO:1071; polynucleotide SEQIDNO:1072 encoding the variable light chain sequence of SEQIDNO:1062; the complementarity determining region (SEQIDNO:1054) encoding the heavy chain sequence of SEQIDNO:1041 or the variable heavy chain sequence of SEQIDNO:1042 , SEQIDNO:1056 and SEQIDNO:1058) polynucleotides; encoding the light chain sequence of SEQIDNO:1061 or the complementarity determining region (SEQIDNO:1074, SEQIDNO:1076 and SEQIDNO:1078) of the variable light chain sequence of SEQIDNO:1062 Polynucleotides; polynucleotides encoding the heavy chain sequence of SEQ ID NO: 1041 or the framework region (SEQ ID NO: 1053, SEQ ID NO: 1055, SEQ ID NO: 1057 and SEQ ID NO: 1059) of the variable heavy chain sequence of SEQ ID NO: 1042; and encoding SEQ ID NO A polynucleotide of the light chain sequence of: 1061 or the framework region (SEQ ID NO: 1073, SEQ ID NO: 1075, SEQ ID NO: 1077 and SEQ ID NO: 1079) of the variable light chain sequence of SEQ ID NO: 1062.

In a preferred embodiment of the present invention, the polynucleotide of the present invention comprises or consists of a polynucleotide encoding a Fab (Fragment Antigen-Binding) fragment having binding specificity for HGF. With respect to antibody Ab28, the polynucleotide encoding the full-length Ab28 antibody comprises or consists of the polynucleotide SEQ ID NO: 1051 encoding the heavy chain sequence of SEQ ID NO: 1041 and the polynucleotide SEQ ID NO: 1061 encoding the light chain sequence: 1071.

In one embodiment, the present invention relates to an isolated polynucleotide comprising a polynucleotide encoding an anti- _{HGFV H antibody} amino acid sequence selected from the group consisting of SEQ ID NO:12, SEQ ID NO:52, SEQ ID NO: 92, SEQ ID NO: 132, SEQ ID NO: 172, SEQ ID NO: 212, SEQ ID NO: 252, SEQ ID NO: 292, SEQ ID NO: 332, SEQ ID NO: 372, SEQ ID NO: 412, SEQ ID NO: 452, SEQ ID NO: 492, SEQ ID NO: 532, SEQ ID NO: 572, or code The polynucleotide of its variant, wherein at least one framework residue (FR residue) has been present in the amino acid substitution or conservative amino acid substitution at the corresponding position in the VH polypeptide of the rabbit anti-HGF antibody.

In another embodiment, the present invention relates to an isolated polynucleotide comprising a polynucleotide sequence encoding an anti- _{HGFV L antibody} amino acid selected from the group consisting of SEQ ID NO:32, SEQ ID NO:72, SEQ ID NO: 112, SEQ ID NO: 152, SEQ ID NO: 192, SEQ ID NO: 232, SEQ ID NO: 272, SEQ ID NO: 312, SEQ ID NO: 352, SEQ ID NO: 392, SEQ ID NO: 432, SEQ ID NO: 472, SEQ ID NO: 512, SEQ ID NO: 552, SEQ ID NO: 592, SEQ ID NO: 632, SEQ ID NO: 672, SEQ ID NO: 712, SEQ ID NO: 752, SEQ ID NO: 792, SEQ ID NO: 832, SEQ ID NO: 872, SEQ ID NO: 912, SEQ ID NO: 952, SEQ ID NO: 992, SEQ ID NO: 1032 or SEQ ID NO: 1072; or code The polynucleotide sequence of its variant, wherein at least one framework residue (FR residue) has been replaced by amino acid substitution or conservative amino acid substitution at the corresponding position in the rabbit anti-HGF antibody VL polypeptide.

In another embodiment, the present invention relates to one or more heterologous polynucleotides comprising sequences encoding polypeptides contained in: SEQ ID NO: 2 and SEQ ID NO: 22, SEQ ID NO: 42 and SEQ ID NO: 62, SEQ ID NO :82 and SEQ ID NO: 102, SEQ ID NO: 122 and SEQ ID NO: 142, SEQ ID NO: 162 and SEQ ID NO: 182, SEQ ID NO: 202 and SEQ ID NO: 222, SEQ ID NO: 242 and SEQ ID NO: 262, SEQ ID NO: 282 and SEQ ID NO: 302, SEQ ID NO: 322 and SEQ ID NO:342, SEQ ID NO:362 and SEQ ID NO:382, SEQ ID NO:402 and SEQ ID NO:422, SEQ ID NO:442 and SEQ ID NO:462, SEQ ID NO:482 and SEQ ID NO:502, SEQ ID NO:522 and SEQ ID NO:542, SEQ ID NO:562 and SEQ ID NO : 582, SEQ ID NO: 602 and SEQ ID NO: 622, SEQ ID NO: 642 and SEQ ID NO: 662, SEQ ID NO: 682 and SEQ ID NO: 702, SEQ ID NO: 722 and SEQ ID NO: 742, SEQ ID NO: 762 and SEQ ID NO: 782, SEQ ID NO: 802 and SEQ ID NO: 822 , SEQ ID NO:842 and SEQ ID NO:862, SEQ ID NO:882 and SEQ ID NO:902, SEQ ID NO:922 and SEQ ID NO:942, SEQ ID NO:962 and SEQ ID NO:982, SEQ ID NO:1002 and SEQ ID NO:1022, and SEQ ID NO:1042 and SEQ ID NO:1062.

In another embodiment, the invention relates to an isolated polynucleotide expressing a polypeptide comprising at least one CDR polypeptide derived from an anti-HGF antibody according to the invention, such as Ab1, Ab2, Ab7 , Ab8, Ab9, Ab10, Ab12, Ab14, Ab19, Ab21, Ab23, Ab24, Ab25 and Ab28, wherein said expressed polypeptide specifically binds to HGF alone or when combined with a polypeptide containing at least one CDR derived from the following anti-HGF antibody Another polynucleotide sequence of the polypeptide specifically binds HGF when it is jointly expressed, wherein said at least one CDR is selected from those CDRs contained in the following VL or VH polypeptides: SEQ ID NO: 2, 22, 42, 62, 82, 102, 122, 142, 162, 182, 202, 222, 242, 262, 282, 302, 322, 342, 362, 382, 402, 422, 442, 462, 482, 502, 522, 542, 562, 582, 602, 622, 642, 662, 682, 702, 722, 742, 762, 782, 802, 822, 842, 862, 882, 902, 922, 942, 962, 982, 1002, 1022, 1042, and 1062. More specifically, said at least one CDR comprises SEQ ID NO: 4, 6, 8, 24, 26, 28, 44, 46, 48, 64, 66, 68, 84, 86, 88, 104, 106, 108, 124 ,126,128,144,146,148,164,166,168,184,186,188,204,206,208,224,226,228,244,246,248,264,266,268,284,286 ,288,304,306,308,324,326,328,344,346,348,364,366,368,384,386,388,404,406,408,424,426,428,444,446,448 ,464,466,468,484,486,488,504,506,508,524,526,528,544,546,548,564,566,568,584,586,588,604,606,608,624 ,626,628,644,646,648,664,666,668,684,686,688,704,706,708,724,726,728,744,746,748,764,766,768,784,786 ,788,804,806,808,824,826,828,844,846,848,864,866,884,886,888,904,906,908,924,926,928,944,946 or 948,964 , 966, 968, 984, 986 or 988, 1004, 1006, 1008, 1024, 1026 or 1028, 1044, 1046, 1048, 1064, 1066 or 1068.

Host cells and vectors comprising the polynucleotides are also contemplated.

The invention also encompasses vectors comprising polynucleotide sequences encoding the variable heavy and light chain polypeptide sequences and individual complementarity determining regions (CDRs or hypervariable regions) as set forth herein, and host cells comprising said vector sequences . In one embodiment of the invention, the host cell is a yeast cell. In another embodiment of the invention, the yeast host cell belongs to the genus Pichia.

Anti-HGF activity

The anti-HGF antibodies and fragments thereof of the present invention having binding specificity for HGF can also be described by their binding strength or affinity for HGF. In one embodiment of the present invention, the anti-HGF antibodies and fragments thereof of the present invention having binding specificity for HGF bind to HGF with a dissociation constant (KD) less than or equal to: 5×10 ⁻⁷ , 10 ^{− 7} , 5×10 ^-8 , 10 ^-8 , 5×10 ^-9 , 10 ^-9 , 5×10 ^-10 , 10 ^-10 , 5×10 ^-11 , 10 ^-11 , 5×10 ^-12 , 10- 12, 5×10 ⁻¹³ , 10 ⁻¹³ , 5×10 −14 , 10 ⁻¹⁴ , 5×10 ⁻¹⁵ or 10 ⁻¹⁵ .

Preferably, anti-HGF antibodies and fragments thereof bind HGF with a dissociation constant less than or equal to 5 x 10 ^-10 . In another embodiment of the invention, the anti-HGF antibodies and fragments thereof of the invention having binding specificity for HGF bind to a linear or conformational HGF epitope.

In another embodiment of the present invention, the anti-HGF antibodies and fragments thereof with binding specificity for HGF of the present invention are less than or equal to 10 ^-4 S ^-1 , 5×10 ^-5 S ^-1 , 10 ^-5 The off-rates of S ⁻¹ , 5×10 ⁻⁶ S ⁻¹ , 10 ⁻⁶ S ⁻¹ , 5×10 ⁻⁷ S ⁻¹ , or 10 ⁻⁷ S ⁻¹ bound to HGF.

In another embodiment of the invention, the anti-HGF antibodies and fragments thereof of the invention having binding specificity for HGF (including those antibodies and fragments thereof having the binding affinities or dissociation constants described above) inhibit or Blocking at least one HGF-related biological activity. The term "HGF biological activity" as used herein refers to any mitogenic, promotile or morphogenic activity of HGF or any activity that occurs as a result of HGF binding to the HGF receptor. In particular, the antibodies and antibody fragments of the invention can be used to inhibit or block HGF-related cell proliferation, invasion, dispersion, metastasis, angiogenesis, fibrosis, and activation of C-met or HGF receptors.

The term "HGF receptor or c-met activation" refers to HGF receptor dimerization or HGF receptor induced tyrosine kinase activity. HGF receptor activation can occur as a result of HGF binding to the HGF receptor, but alternatively can occur independent of any HGF binding to the HGF receptor. HGF biological activity can eg be determined in an in vitro or in vivo hepatocyte growth promoting assay, or more preferably such as the protocol disclosed in Example 12 below, using 4mBr-5 cells. Adult rat hepatocytes in primary culture can be used to test the effect of HGF on hepatocyte proliferation. Alternatively, the effect of an HGF antibody antagonist according to the invention can be determined in an assay suitable for testing the ability of HGF to induce DNA synthesis in other types of cells expressing HGF receptors, such as mink lung cells or human mammary epithelial cells and as described in the Examples below. DNA synthesis can be analyzed, for example, by measuring the incorporation of3H-thymidine into DNA. The effectiveness of HGF receptor antagonists can be measured by their ability to block the proliferation and incorporation of3H-thymidine into DNA. The effect of HGF antagonists can also be tested in vivo in animal models, especially the tumor xenograft models disclosed below. Furthermore, the effect of an anti-HGF antibody or fragment according to the invention on c-met activation can be assessed such as described in the Examples below.

In another embodiment of the present invention, the anti-HGF activity of the anti-HGF antibodies and fragments thereof with binding specificity for HGF of the present invention exhibits the prevention, improvement or alleviation of symptoms of diseases and disorders associated with HGF or the treatment of Anti-HGF activity in the diseases and disorders mentioned above. Non-limiting examples of diseases and disorders associated with HGF are set forth below.

B cell screening and isolation

In one embodiment, the present invention encompasses the preparation and isolation of a clonal population of antigen-specific B cells that can be used to isolate at least one HGF antigen-specific cell that can be used to generate HGF-directed cells with specificity for a desired HGF antigen. Monoclonal antibodies, or nucleic acid sequences corresponding to such antibodies. Methods for the preparation and isolation of such antigen-specific B cell clonal populations are taught, for example, in US Patent Publication No. US2007/0269868 to Carvalho-Jensen et al., the disclosure of which is incorporated herein by reference in its entirety. Methods of making and isolating said clonal populations of antigen-specific B cells are also taught in the Examples herein. Methods of "enriching" cell populations by size or density are known in the art. See, eg, US Patent 5,627,052. These steps can be used in addition to enriching the cell population by antigen specificity.

Methods of Humanizing Antibodies

In another embodiment, the invention encompasses methods for humanizing antibody heavy and light chains. Methods for humanizing antibody heavy and light chains applicable to anti-HGF antibodies are taught, for example, in U.S. Patent Application Publication No. US2009/0022659 to Olson et al. and U.S. Pat. Patent No. 7,935,340, the disclosures of each of which are incorporated herein by reference in their entirety.

Methods for producing antibodies and fragments thereof

In another embodiment, the present invention encompasses methods of producing anti-HGF antibodies and fragments thereof. Methods for the production of anti-HGF antibodies and fragments thereof secreted from polyploid, preferably diploid or tetraploid strains of mating-competent yeast are taught, for example, in U.S. Patent Application Publication No. US2009/0022659 to Olson et al. Neutralization in US Patent No. 7,935,340 to Garcia-Martinez et al., the disclosures of each of which are incorporated herein by reference in their entirety. A preferred yeast for making antibodies is Pichia, and more preferably Pichia pastoris. However, antibodies according to the present invention may also be produced in other yeasts, such as other mating-competent yeasts of the family Saccharomyces, which include the genera Aspenia; ; Dekkeryas; Pseudomyces; Issa; Kazachstania; Kluyveromyces; genus; Tetrasporum; Torulaspora; Pseudomonas; and Zygomyces. Other types of yeast that may be suitable for the preparation of antibody proteins according to the invention include Yarrowia; Rhodosporidium; Candida; Hansenula; Saccharomyces sp;

Other methods of generating antibodies are known to those of ordinary skill. For example, methods for the generation of chimeric antibodies are now well known in the art (see, e.g., U.S. Patent No. 4,816,567 to Cabilly et al; Morrison et al, P.N.A.S. USA, 81:8651-55 (1984); Neuberger, M.S. et al., Nature, 314:268-270 (1985); Boulianne, G.L. et al., Nature, 312:643-46 (1984), the disclosures of which are each incorporated herein by reference in their entirety) .

Similarly, other methods of producing humanized antibodies are now well known in the art (see, e.g., US Patent Nos. 5,530,101, 5,585,089, 5,693,762, and 6,180,370 to Queen et al. ; U.S. Patents 5,225,539 and 6,548,640 to Winter; U.S. Patents 6,054,297, 6,407,213 and 6,639,055 to Carter et al; U.S. Patent 6,632,927 to Adair ; Jones, P.T. et al., Nature, 321:522-525 (1986); Reichmann, L. et al., Nature, 332:323-327 (1988); Verhoeyen, M et al., Science, 239:1534-36 ( 1988), the disclosures of each of which are incorporated herein by reference in their entirety).

The antibody polypeptide having HGF binding specificity of the present invention can also be produced by using conventional techniques well known to those of ordinary skill to construct an expression vector containing an operon encoding an antibody heavy chain and a DNA sequence, wherein the DNA encoding the CDRs required for antibody specificity The sequences are derived from non-human cell sources, preferably rabbit B cell sources, while the DNA sequences encoding the remainder of the antibody chains are derived from human cell sources.

A second expression vector is produced using the same conventional means well known to those of ordinary skill, said expression vector comprising an operon and a DNA sequence encoding the light chain of the antibody, wherein the DNA sequence encoding the CDRs required for antibody specificity is derived from a non-human cell source, Rabbit B cell origin is preferred, while the DNA sequences encoding the remainder of the antibody chains are of human cell origin.

The expression vector is transfected into host cells by conventional techniques well known to those of ordinary skill, thereby producing transfected host cells, which are cultured by conventional techniques well known to those of ordinary skill to produce the antibody polypeptide .

Host cells can be co-transfected with the two expression vectors described above, the first containing DNA encoding the operon and the light chain-derived polypeptide and the second containing the DNA encoding the operon and the heavy chain-derived polypeptide. The two vectors contain different selectable markers, but preferably achieve substantially the same expression of heavy and light chain polypeptides. Alternatively, a single vector can be used that includes DNA encoding both heavy and light chain polypeptides. The coding sequences for the heavy and light chains may comprise cDNA, genomic DNA, or both.

Host cells that may be useful for expressing antibody polypeptides of the invention may include bacterial cells, such as E. coli; or eukaryotic cells, such as Pichia pastoris; other yeast cells; fungal, insect cells; mammalian cells and plant cells. In one embodiment of the invention, a well-defined type of mammalian cell for this purpose may be, for example, a myeloma cell, a Chinese Hamster Ovary (CHO) cell line, an NSO cell line or a HEK293 cell line.

The general methods by which vectors may be constructed, the transfection methods required to produce host cells, and the culture methods required to produce antibody polypeptides from said host cells all include conventional techniques. While the cell line used to produce the antibody is preferably a mammalian cell line, any other suitable cell line may alternatively be used, such as a bacterial cell line, such as a strain derived from E. coli; or a yeast cell line.

Similarly, antibody polypeptides once produced can be purified according to standard procedures in the art, such as cross-flow filtration, ammonium sulfate precipitation, affinity column chromatography, and the like.

The antibody polypeptides described herein can also be used to design and synthesize peptide or non-peptide mimetics that would be suitable for the same therapeutic applications as the antibody polypeptides of the invention. See, eg, Saragobi et al., Science, 253:792-795 (1991), the contents of which are incorporated herein by reference in their entirety.

screening assay

The invention also includes screening assays designed to aid in the identification of HGF-associated diseases and disorders in patients exhibiting symptoms of HGF-associated diseases or disorders.

In some embodiments, antibodies are used as diagnostic tools. Antibodies can be used to determine the amount of HGF present in a sample and/or subject. As will be appreciated by those skilled in the art, such antibodies need not be neutralizing antibodies. In some embodiments, the diagnostic antibody is not a neutralizing antibody. In some embodiments, the diagnostic antibody binds to a different epitope than the neutralizing antibody binds. In some embodiments, the two antibodies do not compete with each other.

In some embodiments, the antibodies disclosed herein are used or provided in assay kits and/or methods for detecting HGF in mammalian tissues or cells, so as to screen/diagnose diseases or disorders associated with changes in HGF levels. The kit includes an antibody that binds HGF and means for demonstrating binding of the antibody to HGF (if present) and, optionally, the amount of HGF protein present. Various constructs for indicating the presence of antibodies can be used. For example, fluorophores, other molecular probes, or enzymes can be attached to antibodies and the presence of antibodies can be observed in various ways. Screening methods for such disorders may involve using a kit, or simply using one of the disclosed antibodies and determining whether the antibody binds to HGF in a sample. As will be appreciated by those skilled in the art, high or higher levels of HGF will result in a greater amount of antibody binding to HGF in the sample. Therefore, the degree of antibody binding can be used to determine how much HGF is in the sample. A subject or sample having an amount of HGF greater than a predetermined amount (eg, the amount or variation that would be present in a person not suffering from an HGF-related disorder) can be characterized as having an HGF-mediated disorder. In some embodiments, the antibody is administered to a subject on a statin in order to determine whether the statin affects the amount of HGF in the subject.

The present invention also relates to an in vivo imaging method for detecting the presence of cells expressing HGF, said method comprising administering a diagnostically effective amount of a diagnostic composition. Such in vivo imaging is useful, for example, for detecting or imaging HGF-expressing cells or organs, and may be useful as part of a planning protocol for designing effective cancer treatment regimens. Treatment regimens may include, for example, one or more of radiation, chemotherapy, cytokine therapy, gene therapy, and antibody therapy, as well as anti-HGF antibodies or fragments thereof.

The present invention further provides a kit for detecting the binding of the anti-HGF antibody of the present invention to HGF. In particular, the kits can be used to detect the presence of HGF or immunologically active fragments thereof that are specifically reactive with the anti-HGF antibodies of the invention. The kit may also include an antibody that binds to a substrate, a secondary antibody reactive with the antigen, and reagents for detecting the reaction of the secondary antibody with the antigen. Such kits may be ELISA kits and may include substrates, primary and secondary antibodies (where appropriate) and any other required reagents, such as detectable moieties, enzyme substrates and color reagents as described herein. The diagnostic kit can also be in the form of an immunoblotting kit. The diagnostic kit can also be in the form of a chemiluminescence kit (MesoScale Discovery, Gaithersburg, MD). The diagnostic kit may also be a lanthanide-based detection kit (PerkinElmer, San Jose, CA).

The skilled clinician will appreciate that biological samples include, but are not limited to, serum, plasma, urine, saliva, mucus, pleural fluid, synovial fluid, and spinal fluid.

Methods of improving or alleviating the symptoms of, or treating or preventing, diseases and disorders associated with HGF and/or HGF/HGFR (c-met) interaction and/or c-met activation

The anti-HGF antibodies or fragments thereof described herein are more suitable for ameliorating or alleviating the symptoms of, or treating or preventing, diseases and disorders associated with HGF, including those associated with HGF, based on their binding and functional properties. HGF-R interactions are associated with those diseases and disorders associated with HGF-associated activation of c-met. Anti-HGF antibodies or fragments thereof and combinations described herein can also be administered in therapeutically effective amounts in the form of pharmaceutical compositions described in more detail below to patients in need of treatment of diseases and disorders associated with HGF.

In one embodiment of the invention, the anti-HGF antibodies or fragments thereof described herein are suitable for ameliorating or alleviating the symptoms of, or treating or preventing, the following non-limiting list of diseases and disorders: cancer, including ovarian Cancer, breast cancer, lung cancer (small cell or non-small cell), colon and colorectal cancer, prostate cancer, pancreatic cancer, kidney cancer, stomach cancer, liver cancer, bladder cancer, thyroid cancer, endometrial cancer, head and neck cancer, Melanoma, sarcoma, leukemia; lymphoma; and brain tumors (eg, glioblastoma) in children or adults; macular degeneration; Alzheimer's disease; and malaria infection. In a preferred embodiment, the disease is selected from cancer or macular degeneration.

In another embodiment of the present invention, the present invention provides the use of the anti-HGF antibody of the present invention in the manufacture of a medicament for the treatment and/or prophylaxis of diseases such as cancer, tumors, cell proliferative disorders, immune (such as autoimmune) disorders and/or angiogenesis-related disorders. In another embodiment of the present invention, the present invention provides the use of the nucleic acid of the present invention in the preparation of a medicament for the treatment and/or prophylaxis of diseases such as cancer, tumors, cell proliferative disorders, immune (such as autoimmune) disorders and/or angiogenesis-related disorders.

In another embodiment of the present invention, the present invention provides the use of the expression vector of the present invention in the preparation of medicaments for the treatment and/or prophylaxis of diseases such as cancer, tumors, cell proliferative disorders, immune ( such as autoimmune) disorders and/or angiogenesis-related disorders. In another embodiment of the present invention, the present invention provides the use of the host cell of the present invention in the preparation of a medicament for the treatment and/or prophylaxis of diseases such as cancer, tumors, cell proliferative disorders, immune ( such as autoimmune) disorders and/or angiogenesis-related disorders. In another embodiment of the invention, the invention provides the use of the preparation of the invention for the manufacture of a medicament for the treatment and/or prophylaxis of diseases such as cancer, tumors, cell proliferative disorders, immune (such as autoimmune) disorders and/or angiogenesis-related disorders.

In another aspect, the present invention provides the use of the kit of the present invention in the manufacture of a medicament for the treatment and/or prophylaxis of diseases such as cancer, tumors, cell proliferative disorders, immune (such as autoimmune ) disorders and/or angiogenesis-related disorders.

In a preferred embodiment of the invention, the invention provides methods and compositions useful for modulating conditions associated with dysregulation of the HGF/c-met signaling axis. The HGF/c-met signaling pathway is involved in a variety of biological and physiological functions including, for example, cell proliferation and angiogenesis. Accordingly, in one aspect, the invention provides a method comprising administering to a subject an antibody of the invention. In one aspect, the invention provides a method of inhibiting c-met activated cell proliferation; said method comprising contacting a cell or tissue with an effective amount of an antibody of the invention, whereby cell proliferation associated with c-met activation is inhibited. In another aspect, the invention provides a method of treating a pathological condition associated with dysregulation of c-met activation in a subject, the method comprising administering to the subject an effective amount of an antibody of the invention, whereby This treats the condition. In another aspect, the invention provides a method of inhibiting the growth of cells expressing c-met or hepatocyte growth factor, or both, comprising contacting said cells with an antibody of the invention, thereby causing said Inhibition of cell growth. In one embodiment, the cells are contacted (eg, via paracrine action) with HGF expressed by a different cell. In another aspect, the invention provides a method of therapeutically treating a mammal having a cancerous tumor comprising cells expressing c-met or hepatocyte growth factor, or both, comprising administering to said mammal The animal is administered an effective amount of an antibody of the invention, thereby effectively treating the mammal. In one embodiment, the cells are contacted (eg, via paracrine action) with HGF expressed by a different cell. In another aspect, the present invention provides a method for treating or preventing a cell proliferative disorder associated with increased expression or activity of c-met or hepatocyte growth, or both, comprising requesting a patient in need of such treatment A subject is administered an effective amount of an antibody of the invention, thereby effectively treating or preventing said cell proliferative disorder. In one embodiment, the proliferative disorder is cancer. In another aspect, the present invention provides a method for inhibiting the growth of a cell, wherein the growth of the cell depends at least in part on the growth enhancing effect of c-met or hepatocyte growth factor or both, the method comprising The cells are contacted with an effective amount of an antibody of the invention, whereby growth of the cells is inhibited. In one embodiment, the cells are contacted (eg, via paracrine action) with HGF expressed by a different cell.

In another aspect, the present invention provides a method of therapeutically treating a tumor in a mammal, wherein the growth of said tumor depends at least in part on the growth enhancing effect of c-met or hepatocyte growth factor or both, The method comprises contacting the cell with an effective amount of an antibody of the invention, thereby effectively treating the tumor. In one embodiment, the cells are contacted (eg, via paracrine action) with HGF expressed by a different cell.

The methods of the invention can be used to treat any suitable pathological condition in which antagonism of HGF or the interaction of HGF/c-met is therapeutically advantageous, such as cells and/or tissues associated with dysregulation of the HGF/c-met signaling pathway . In one embodiment, the cells targeted by the methods of the invention are cancer cells. For example, the cancer cell may be a cell selected from the group consisting of: breast cancer cells, colorectal cancer cells, lung cancer cells, papillary cancer cells (such as papillary cancer cells of the thyroid), colon cancer cells, pancreatic cancer cells , ovarian cancer cells, cervical cancer cells, central nervous system cancer cells, osteogenic sarcoma cells, kidney cancer cells, liver cell cancer cells, bladder cancer cells, gastric cancer cells, head and neck squamous cell carcinoma cells, melanoma cells and leukemia cells. In one embodiment, the cells targeted by the methods of the invention are hyperproliferative and/or hyperplastic cells. In one embodiment, the cells targeted by the methods of the invention are dysplastic cells. In another embodiment, the cells targeted by the methods of the invention are metastatic cells.

The methods of the invention may further comprise additional treatment steps. For example, in one embodiment, a method further comprises the step of exposing the targeted cells and/or tissues (eg, cancer cells) to radiation therapy or a chemotherapeutic agent.

As noted, c-met activation is an important biological process whose dysregulation contributes to numerous pathological conditions. Thus, in one embodiment of the methods of the invention, the targeted cells (eg, cancer cells) are cells that have enhanced activation of c-met compared to normal cells of the same tissue origin. In one embodiment, the methods of the invention result in the death of the targeted cell. For example, exposure to an antagonist of the invention can render cells incapable of signaling in the c-met pathway, resulting in cell death.

Dysregulation of c-met activation (and thus signaling) can result from a variety of cellular changes including, for example, overexpression of HGF (c-met's cognate ligand) and/or c-met itself. Thus, in some embodiments, the methods of the invention comprise targeting a cell wherein c-met or hepatocyte growth factor or both are more mediated by the cell (e.g., a cancer cell) than a normal cell of the same tissue origin. Fully express. Cells expressing c-met can be regulated by HGF from various sources, ie in an autocrine or paracrine manner. For example, in one embodiment of the methods of the invention, the targeted cells are contacted/bound by hepatocyte growth factor expressed (eg via paracrine action) in a different cell. The different cells may be of the same or different tissue origin. In one embodiment, the targeted cell is contacted/bound by HGF expressed by the targeted cell itself (eg, via an autocrine action/loop). C-met activation and/or signaling can also occur independent of ligand. Thus, in one embodiment of the methods of the invention, activation of c-met in the targeted cell occurs independently of the ligand.

Conditions treated with anti-HGF antibodies or fragments of the invention include any condition that would benefit from treatment with an anti-HGF antibody or fragment or method of use thereof according to the invention. Such conditions include chronic and acute conditions or diseases, including those pathological conditions that predispose a mammal to the condition in question. Non-limiting examples of conditions to be treated herein include malignant and benign tumors; non-leukemic and lymphoid malignancies; neuronal, glial, astrocyte, hypothalamic and other glandular, macrophage, epithelial Cellular, stromal, and blastocoel disorders; and inflammatory, immune, and other angiogenesis-related disorders. As noted previously, cell proliferative disorders treatable in accordance with the present invention include disorders associated with some degree of abnormal cell proliferation. In one embodiment, the cell proliferative disorder is cancer.

Specific examples of the cancer include squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, peritoneal carcinoma, hepatocellular carcinoma, gastrointestinal cancer, pancreatic cancer, glioblastoma Cytoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatocellular tumor, breast cancer, colon cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney cancer, liver cancer, prostate cancer, vulva Cancer, thyroid cancer, liver cancer and various types of head and neck cancer.

Another class of disorders that can be treated with the anti-HGF antibodies or fragments of the invention include those involving dysregulation of angiogenesis. These conditions include both non-neoplastic and neoplastic conditions. Neoplastic conditions include, but are not limited to, those cancers described above. Non-neoplastic conditions include, but are not limited to, undesirable or abnormal hypertrophy, arthritis, rheumatoid arthritis (RA), psoriasis, psoriatic plaques, sarcoidosis, atherosclerosis, atherosclerosis Diabetic and other proliferative retinopathy, including retinopathy of prematurity, retrolentic fibroplasia, neovascular glaucoma, age-related macular degeneration, diabetic macular edema, corneal neovascularization, corneal transplantation neovascularization , corneal graft rejection, retinal/choroidal neovascularization, canthal neovascularization (iris redness), ocular neovascular disease, vascular restenosis, arteriovenous malformation (AVM), meningioma, hemangioma, angiofibroma , thyroid hyperplasia (including Graves' disease), corneal and other tissue transplantation, chronic inflammation, pulmonary inflammation, acute lung injury/ARDS, sepsis, essential pulmonary hypertension, malignant pulmonary effusion, cerebral edema (eg, with acute stroke/closed head injury/trauma-related), synovial inflammation, pannus formation in RA, myositis ossificans, hypertrophic bone formation, osteoarthritis (OA), refractory ascites, polycystic Ovarian disease, endometriosis, 3rd compartment humoral disease (pancreatitis, compartment syndrome, burns, enteropathy), uterine fibroids, premature labor, chronic inflammation such as IBD (Crohn's disease and ulcerative colon inflammation), renal allograft rejection, inflammatory bowel disease, nephrotic syndrome, undesired or abnormal mass growth of tissue (not cancer), haemophilic joints, hypertrophic scarring, inhibited hair growth, Osler- Weber syndrome, pyogenic granulomatoma retrolentic fibroplasia, scleroderma, trachoma, vascular adhesions, synovitis, dermatitis, preeclampsia, ascites, pericardial effusions (such as those associated with pericarditis ) and pleural effusion.

In preferred embodiments, antibodies to HGF are used with one or more other therapeutic agents or regimens for the treatment of various cancers. In certain embodiments, antibodies to HGF are used with one or more specific therapeutic agents to treat or prevent malaria. In certain embodiments, antibodies specific for HGF are used with one or more specific therapeutic agents to treat or prevent proliferative diabetic retinopathy. In certain embodiments, two, three, or more agents may be administered, given the condition and degree of treatment desired. In certain embodiments, the agents may be provided together by inclusion in the same formulation. In certain embodiments, the agent and the specific binding agent to HGF may be provided together by inclusion in the same formulation. In certain embodiments, the agents may be formulated separately and provided together by inclusion in a therapeutic kit. In certain embodiments, the agent and the specific binding agent to HGF may be formulated separately and provided together by inclusion in a therapeutic kit. In certain embodiments, the agents may be provided separately.

The methods of the invention may be useful in the amelioration and/or treatment of diseases or conditions involving abnormal angiogenesis. "Aberrant angiogenesis" as used herein refers to angiogenesis that does not occur during normal biological processes such as development, reproduction, wound healing, and the like. In some embodiments, angiogenesis that can be reduced using the methods of the invention can be stimulated by a factor selected from the group consisting of: vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), lipopolysaccharide (LPS ), epidermal growth factor (EGF), interleukin-6 (IL-6), platelet-derived growth factor (PDGF), tumor necrosis factor (TNFalpha), hepatocyte growth factor (HGF), and combinations thereof.

Because angiogenesis is involved in various pathological processes, the methods of the present invention are applicable to the treatment and/or amelioration of diseases such as cancer (including metastatic cancer), ocular neovascularization (such as macular degeneration), inflammatory diseases (such as arthritis )Wait.

Tumors often depend on the formation of new blood vessels for continued growth and/or the development of metastases. Therefore, the method of the present invention is applicable to the amelioration and/or treatment of tumors. A subject to whom a chlorotoxin agent may be administered may have a tumor that has metastasized or that has metastasized. A subject may have one or more cancer metastases. In some embodiments, the tumor and/or metastases are reduced in size.

In some embodiments, the subject suffers from or is at risk of developing a condition or disease characterized by choroidal neovascularization. Such conditions include, but are not limited to, macular degeneration, myopia, ocular trauma, pseudoxanthoma elasticum, and combinations thereof.

Macular degeneration is the leading cause of vision loss and blindness in Americans 65 and older. Macular degeneration usually occurs in an age-related form (often called AMD or ARMD), but juvenile macular degeneration can also occur. In AMD/ARMD, the macula is the part of the retina responsible for the dramatic deterioration of central vision. Macular degeneration is usually diagnosed as dry (non-neovascular) or wet (neovascular). In dry macular degeneration, yellowish spots called drusen begin to accumulate from deposits or debris of degenerated tissue that mostly surrounds the macula. Central vision usually occurs less gradually and is not as severe as vision loss in wet macular degeneration.

As the name "neovascular" suggests, wet macular degeneration is characterized by abnormal growth of new blood vessels, such as in the macula. These new blood vessels can grow under the retina, leaking blood and body fluids. Such leakage results in permanent damage to the light-sensitive retinal cells, which die and create blind spots in central vision. Wet macular degeneration can be further divided into two categories. In the recessive form of wet macular degeneration, new blood vessels growing under the retina are less prominent and less leaky, usually producing less severe vision loss. In the typical form of wet macular degeneration, vascular growth and scarring have very clear, delineated outlines that can be visualized under the retina. Classic wet macular degeneration is also known as classic choroidal neovascularization and often results in more severe vision loss.

Given the role of angiogenesis in wet macular degeneration, which encompasses many AMD/ARMD conditions, the methods of the present invention are applicable to the treatment and/or amelioration of such conditions. Current therapies for wet macular degeneration involve angiogenesis inhibitors, such as Lucentis™, Macugen™ and/or Visudyne™, optionally in combination with photodynamic therapy (PDT) to target drugs to specific cells. Photocoagulation is also used to treat wet macular degeneration, in which a higher energy laser beam is used to create small burns in areas of the retina with abnormal blood vessels.

In some embodiments, the subject suffers from wet macular degeneration and/or age-related macular degeneration. In subjects suffering from wet macular degeneration, the subject may suffer from recessive or classic forms. In some embodiments, the chlorotoxin agent causes regression of existing neovascularization. In some embodiments, the chlorotoxin agent prevents neovascularization. In certain embodiments, chlorotoxin agents are combined with other treatments for wet macular degeneration, such as photocoagulation, treatment with other angiogenesis inhibitors, photodynamic therapy, and the like.

The above are intended to be exemplary diseases and conditions in which administration of an anti-HGF antibody or fragment according to the invention may be therapeutically beneficial.

apply

In one embodiment of the invention, the anti-HGF antibodies or HGF-binding fragments thereof described herein and combinations of said antibodies or antibody fragments are administered to Subject administration. In a preferred embodiment of the invention, the anti-HGF antibodies or HGF-binding fragments thereof described herein, and combinations of said antibodies or antibody fragments, are administered to a subject at a concentration of about 0.4 mg per kg of the recipient subject's body weight. apply. In a preferred embodiment of the invention, the anti-HGF antibodies or HGF-binding fragments thereof as described herein and combinations of said antibodies or antibody fragments are administered once every twenty-six weeks or less than once every twenty-six weeks, such as every ten Administration is to the recipient subject once every six weeks or less, once every eight weeks or less, or every four weeks or less.

Those of ordinary skill will be able to determine the effective dose and frequency of administration via, for example, routine experimentation guided by the disclosure herein and the teachings in: Goodman, L.S., Gilman, A., Brunton, L.L., Lazo, J.S. and Parker, K.L. (2006). Goodman & Gilman's thepharmacological basis of therapeutics. NewYork: McGraw-Hill; Howland, R.D., Mycek, M.J., Harvey, R.A., Champe, P.C. and Mycek, M.J. (2006). Pharmacology. Lippincott Williams &Wilkins; and Golan, D.E. (2008). Principles of pharmacology: thepathophysiologic basis of drug therapy. Philadelphia, Pa. [et al]: Lippincott Williams & Wilkins.

In another embodiment of the invention, the anti-HGF antibodies or HGF-binding fragments thereof described herein and combinations of said antibodies or antibody fragments are administered to a subject in the form of a pharmaceutical formulation.

"Pharmaceutical composition" means a chemical or biological composition suitable for administration to a mammal. Such compositions may be specially formulated for administration via one or more of a variety of routes including, but not limited to, buccal, epicutaneous, epidural, inhalation, intraarterial, cardiac Intraventricular, intradermal, intramuscular, intranasal, intraocular, intraperitoneal, intraspinal, intrathecal, intravenous, oral, parenteral, rectal via enema or suppository, subcutaneous, subdermal, sublingual , percutaneous and transmucosal. Additionally, administration may be by means of injection, powder, liquid, gel, drops or other means of administration.

In one embodiment of the invention, the anti-HGF antibodies or HGF-binding fragments thereof described herein, and combinations of said antibodies or antibody fragments, may optionally be administered in combination with one or more active agents. Such active agents include analgesics, antipyretics, anti-inflammatory agents, antibiotics, antiviral agents and anticytokine agents. Active agents include TNF-α, IL-2, IL-4, IL-6, IL-10, IL-12, IL-13, IL-18, IFN-α, IFN-γ, BAFF, CXCL13, IP-10 , VEGF, EPO, EGF, HRG, hepatocyte growth factor (HGF), agonists, antagonists and modulators of hepcidin, including antibodies reactive to any of the above, and Any reactive antibody. Active agents also include 2-arylpropionic acid, Aceclofenac, Acemetacin, Acetylsalicylic acid (Aspirin), Alclofenac, Alminoprofen (Alminoprofen), Amoxiprin, Ampyrone, Arylalkanoic Acids, Azapropazone, Benorylate/Benorilate, Benoxaprofen (Benoxaprofen), Bromfenac, Carprofen, Celecoxib, Choline Magnesium Salicylate, Clofezone, COX-2 Inhibitors, Dexibuprofen (Dexibuprofen), Dexketoprofen, Diclofenac, Diflunisal, Droxicam, Ethenzamide, Etodolac, Etoricoxib (Etoricoxib), Faislamine, Fenamic acid, Fenbufen, Fenoprofen, Flufenamic acid, Flunoxaprofen, Fluoride Flurbiprofen, Ibuprofen, Ibuproxam, Indometacin, Indoprofen, Kebuzone, Ketoprofen ( Ketoprofen, Ketorolac, Lornoxicam, Loxoprofen, Lumiracoxib, Magnesium Salicylate, Meclofenamic acid, Meclofenamic acid Mefenamic acid, Meloxicam, Metamizole, Methyl salicylate, Mofebutazone, Nabumetone, Naproxen, N -Arylanthranilic acid, Oxametacin, Oxaprozin, Oxicams, Oxyphenbutazone, Parecoxib, Phenazone ( Phenazone), phenylbutazone, phenylbutazone, piroxicam (Piroxicam), pirprofen (Pirp rofen), profen, proglumetacin, pyrazolidine derivatives, rofecoxib, salicylate, salicylamide, salicylate, sulfinpyrazone , Sulindac, Suprofen, Tenoxicam, Tiaprofenic acid, Tolfenamic acid, Tolmetin, and Valdecoxib. Antibiotics include Amikacin, Aminoglycoside, Amoxicillin, Ampicillin, Ansamycin, Arsphenamine, Azithromycin, Azlocillin, Aztreonam, Bacitracin, Carbacephem, Carbapenem, Carbenicillin, Cefaclor, Cefaclor Cefadroxil, Cefalexin, Cefalothin, Cefalotin, Cefamandole, Cefazolin, Cefdinir, Cefditoren ), Cefepime, Cefixime, Cefoperazone, Cefotaxime, Cefoxitin, Cefpodoxime, Cefprozil , Ceftazidime, Ceftibuten, Ceftizoxime, Ceftobiprole, Ceftriaxone, Cefuroxime, Cephalosporin, Chloramphenicol, Cilastatin, Ciprofloxacin, Clarithromycin, Clindamycin, Cloxacillin, Colistin, Compound New Co-trimoxazole, Dalfopristin, Demeclocycline, Dicloxacillin, Dirithromycin, Doripenem, Doxycycline (Doxycycline), Enoxacin, Ertapenem, Erythromycin, Ethambutol, Flucloxacillin, Fosfomycin, Furazolidone (Furazolidone), fusidic acid, plus Gatifloxacin, Geldanamycin, Gentamicin, Glycopeptides, Herbimycin, Imipenem, Isoniazid , Kanamycin, Levofloxacin, Lincomycin, Linezolid, Lomefloxacin, Loracarbef, Macrolides, Sulfa Mafenide, Meropenem, Meticillin, Metronidazole, Mezlocillin, Minocycline, Monobactams , Moxifloxacin, Mupirocin, Nafcillin, Neomycin, Netilmicin, Nitrofurantoin, Norfloxacin , Ofloxacin, Oxacillin, Oxytetracycline, Paromomycin, Penicillin, Penicillins, Piperacillin, Palymycin ( Platensimycin), Polymyxin B, Polypeptide, Prontosil, Pyrazinamide, Quinolone, Quinupristin, Rifampicin ( Rifampicin), Rifampin, Roxithromycin, Spectinomycin, Streptomycin, Sulfonamide, Sulfamethiadiazole, Sulfanilimide, Sulfasalazine, Sulfisoxazole, Sulfonamides, Teicoplanin, Telithromycin, Tetracycline, Tetracyclines, Ticarcillin, Sulfonamides Metronidazole, Tobramycin, Trimethoprim m), Trimethoprim-Sulfamethoxazole, Troleandomycin, Trovafloxacin and Vancomycin. Active agents also include Aldosterone, Beclometasone, Betamethasone, Corticosteroid, Cortisol, Cortisone acetate, Deoxycorticosterone acetate ), Dexamethasone, Fludrocortisone acetate, Glucocorticoids, Hydrocortisone, Methylprednisolone, Prednisolone, Prednisone, steroids, and triamcinolone. Any suitable combination of these active agents is also contemplated.

A "pharmaceutical excipient" or "pharmaceutically acceptable excipient" is a carrier, usually a liquid, in which an active therapeutic agent is formulated. In one embodiment of the invention, the active therapeutic agent is a humanized antibody or one or more fragments thereof as described herein. Excipients generally do not provide any pharmacological activity to the formulation, but they may provide chemical and/or biological stability and release characteristics. Exemplary formulations can be found, eg, in Remington's Pharmaceutical Sciences, 19th Ed., Grennaro, A. Ed., 1995, which is incorporated herein by reference.

As used herein, "pharmaceutically acceptable carrier" or "excipient" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents that are physiologically compatible . In one embodiment, the carrier is suitable for parenteral administration. Alternatively, the carrier may be suitable for intravenous, intraperitoneal, intramuscular or sublingual administration. Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. The use of such media and agents for pharmaceutically active substances is well known in the art. Unless any conventional media or agents are incompatible with the active compounds, they are contemplated for use in the pharmaceutical compositions of the present invention. Supplementary active compounds can also be incorporated into the compositions.

Pharmaceutical compositions typically must be sterile and stable under the conditions of manufacture and storage. The present invention encompasses pharmaceutical compositions in lyophilized form. The composition can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (eg, glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. The present invention also encompasses the inclusion of stabilizers in the pharmaceutical compositions. Proper fluidity can be maintained, for example, by the use of coatings such as lecithin, by maintenance of the desired particle size (in the case of dispersions), and by the use of surfactants.

In many cases it will be preferable to include isotonic agents, for example sugars, polyalcohols (such as mannitol, sorbitol) or sodium chloride, in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example monostearate salts and gelatin. In addition, basic polypeptides may be formulated in delayed release formulations, eg, in compositions comprising sustained release polymers. The active compounds can be prepared with carriers that will protect the compound against rapid release, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers may be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, polylactic acid and polylactic acids, polyglycolic acid copolymers (PLG). Many methods for the preparation of such formulations are known to those skilled in the art.

With respect to each of the described embodiments, the compounds can be administered via a variety of dosage forms. Any biologically acceptable dosage forms and combinations thereof known to those of ordinary skill in the art are contemplated. Examples of such dosage forms include, but are not limited to, reconstitutable powders, elixirs, liquids, solutions, suspensions, emulsions, powders, granules, granules, microparticles, dispersible granules, cachets, inhalants, aerosol inhalants , patches, particle inhalants, implants, depot implants, injectables (including subcutaneous, intramuscular, intravenous, and intradermal), infusions, and combinations thereof.

The above description of various illustrated embodiments of the invention is not intended to be exhaustive or to limit the invention to the precise forms disclosed. The following examples are presented to further provide a complete disclosure and description of how one of ordinary skill can make and use the invention, and are not intended to limit the scope of the invention. Attempts have been made to ensure accuracy with respect to numbers used (eg amounts, temperature, concentrations, etc.) but some experimental errors and deviations should be allowed for. Unless indicated otherwise, parts are parts by weight; molecular weight is average molecular weight; temperature is in degrees Celsius; and pressure is at or near atmospheric.

The following examples are presented to further provide a complete disclosure and description of how one of ordinary skill can make and use the invention, and are not intended to limit the scope of the invention. Attempts have been made to ensure accuracy with respect to numbers used (eg amounts, temperature, concentrations, etc.) but some experimental errors and deviations should be allowed for. Unless indicated otherwise, parts are parts by weight; molecular weight is average molecular weight; temperature is in degrees Celsius; and pressure is at or near atmospheric.

Example

Example 1 Generation of Concentrated Antigen-Specific B Cell Antibody Cultures

Antibody panels are obtained by immunizing conventional antibody host animals to take advantage of the natural immune response to the target antigen of interest. Typically, the host used for immunization is a rabbit or other host that produces antibodies using a similar maturation process and provides an antibody population that produces antigen-specific B cells with considerable diversity (eg, epitope diversity). Primary antigen immunizations can be performed using complete Freund's adjuvant (CFA), and subsequent boosts are achieved with incomplete adjuvant. About 50-60 days after immunization, preferably at day 55, the antibody titer is tested and if the titer is determined to be suitable, the antibody selection (ABS) process is started. Two key criteria for the initiation of ABS are robust antigen recognition and functional modification activity in polyclonal sera.

Antibody Selection Potency Evaluation

To identify and characterize antibodies that bind to huHGF, antibody-containing solutions were tested by ELISA. Briefly, neutravidin-coated plates (ThermoScientific) were blocked with ELISA buffer (0.1 mg/mL BSA, 1×PBS pH7.4, 0.002% Tween20 and 0.005% sodium azide) at room temperature 1 hour. The plates were then coated with 1 Dg/mL biotinylated huHGF in ELISA buffer for 1 hour at room temperature. This was followed by a washing step (3x with PBS plus 0.05% Tween20) and a second blocking with ELISA buffer. Recombinant antibody was then added to the plate and incubated for 1 hour at room temperature, and then washed 3 times with PBS/Tween solution. For color development, anti-rabbit Fc-HRP (1 :5000 dilution in ELISA buffer) was added to the wells and incubated at room temperature for 45 minutes. After 3 washing steps with PBS/Tween solution, the plate was developed with TMB substrate for 3 minutes, stopped with 0.5M HCl and read at 450 nm.

Upon determination of positive antibody titers, animals were sacrificed and the source of B cells isolated. These sources include: spleen, lymph nodes, bone marrow, and peripheral blood mononuclear cells (PBMC). A single cell suspension is generated and the cell suspension is washed to make it suitable for low temperature long term storage. The cells are then usually frozen.

To begin the antibody identification process, a small aliquot of the frozen cell suspension is thawed, washed, and placed in tissue culture medium. These suspensions were then mixed with a biotinylated form of the antigen used to generate an immune response in the animals, and antigen-specific cells were recovered using the Miltenyi magnetic bead cell selection method. Perform specific enrichment using streptavidin beads. The enriched population is recovered and the next stage of specific B cell isolation proceeds.

Example 2: Generation of Cultures Containing Clonal Antigen-Specific B Cells

Concentrated B cells produced according to Example 1 were then plated in 96-well microtiter plates at varying cell densities per well. Typically, this density is 50, 100, 250 or 500 cells per well in each set of 10 plates. The medium was supplemented with 4% activated rabbit T cell conditioned medium and 50K cryo-irradiated EL4B feeder cells. These cultures were left undisturbed for 5-7 days, at which time supernatants containing secreted antibodies were harvested and assessed for target properties in separate assay settings. The remaining supernatant was left intact and the plates were frozen at -70°C. Under these conditions, the culture process typically results in wells containing a mixed cell population comprising a clonal population of antigen-specific B cells, ie a single well will contain only a single monoclonal antibody specific for the desired antigen.

Example 3: Screening of Monoclonal Antibodies with Desired Specificity and/or Functional Properties in Antibody Supernatants

Antibody-containing supernatants from wells containing clonal antigen-specific B cell populations generated according to Example 2 were first screened for antigen recognition using the ELISA method. This includes selective antigen immobilization (e.g. capture of biotinylated antigen by streptavidin-coated plates), non-specific antigen plate coating, or via antigen accumulation strategies (e.g. selective antigen capture followed by binding partner addition to generate heteromeric protein-antigen complex). Antigen-positive well supernatants are then optionally tested in a functional modification assay that is strictly dependent on the ligand. One such example is an in vitro protein-protein interaction assay that recreates the natural interaction of an antigen ligand with a recombinant receptor protein. Alternatively, a ligand-dependent and easily monitored cell-based response (eg, a proliferative response) is employed. Supernatants showing clear antigen recognition and potency were considered positive wells. Cells derived from initially positive wells then transitioned to the antibody recovery phase.

Example 4: Recovering a Single Antibody-Producing B Cell with a Desired Antigen Specificity

Antigen-specific B cells (generated according to Example 2 or 3) were isolated and used to clone antibody sequences as disclosed in the Examples below. The cells can be used immediately or snap frozen in eppendorf PCR tubes and stored at -80°C until antibody sequence recovery is initiated.

Example 5: Isolation of Antibody Sequences from Antigen-Specific B Cells

Antibody sequences were recovered from single isolated B cells generated according to Example 4 or from antigen-specific B cells isolated from a clonal B cell population obtained according to Example 2 using a combinatorial RT-PCR based approach. Primers are designed to anneal in conserved and constant regions of the immunoglobulin genes of interest (heavy and light chains), such as rabbit immunoglobulin sequences, and a two-step nested PCR recovery step is used to obtain antibody sequences. The amplicons in each well were analyzed for recovery and size integrity. The initial heavy and light chain amplicon fragments were then cloned into expression vectors and transformed into bacteria for plasmid propagation and production. Colonies were selected for sequence characteristics.

Example 6: Recombinant Production of Monoclonal Antibodies with Desired Antigen Specificity and/or Functional Properties

The correct full-length antibody sequence was determined for each well containing a single monoclonal antibody and small-scale purified DNA was prepared. This DNA is then used to transfect mammalian cells to produce recombinant full-length antibodies. Antigen recognition and functional properties of the crude antibody product were tested to confirm the original properties found in the recombinant antibody protein. When appropriate, large-scale transient mammalian transfections were accomplished and antibodies were purified via protein A affinity chromatography. KD is assessed as well as IC50 in potency assays using standard methods such as surface plasmon resonance or biofilm interferometry.

Example 7: Preparation of antibodies binding to HuHGF

By using the antibody selection protocol described herein, a pool of antibodies was generated that included those antibodies that exhibited potent functional antagonism of HGF. The antibodies elucidate various HGF epitopes and thus may provide a useful alternative to or adjunct to antibodies targeting previously identified HGF epitopes.

Example 8: ELISA reactivity of recombinantly expressed anti-HGF antibody of the present invention

To characterize the ability of recombinantly expressed antibodies of the invention to bind to human HGF, antibody-containing solutions were tested by ELISA. All incubations, except for coating, were done at room temperature. Briefly, Immulon4Hbx plates (ThermoScientific) were coated with a solution (1 μg/mL in LPBS) containing HGF (R&D Systems, catalog #294-HGN/CF) overnight at 4°C. HGF coated plates were then washed three times in wash buffer (PBS, 0.05% Tween-20). The plates were then blocked for about one hour using blocking solution (PBS, 0.5% fish skin gelatin). The blocking solution is then removed and the plate is then incubated with the antibody dilution series to be tested for approximately one hour. At the end of this incubation, the plate was washed three times with wash buffer and washed with a solution containing the secondary antibody (peroxidase-conjugated affinity purified F(ab') ₂ fragment goat anti-human IgG, Fc fragment specific ( Jackson Immunoresearch)) were further incubated for about 45 minutes and washed three times. At this time, the substrate solution (TMB peroxidase substrate, BioFx) was incubated in the dark for 3 to 5 minutes. The reaction was stopped by adding a solution containing HCl (0.5 M) and the plate was read in a plate reader at 450 nm.

Antibodies according to the invention (i.e. Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19 determined as described herein , Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28) ELISA reactivity is included in Figures 1-27.

Example 9: Determination of the affinity constant of the recombinantly expressed anti-HGF antibody of the present invention

determination of affinity constant

The binding affinities of Ab8 and Ab10 were determined using Fab fragments. For Ab12, a full length antibody was used. Papain digestion was performed using immobilized papain (Thermo/Pierce) following the manufacturer's instructions. Briefly, purified antibodies were incubated with immobilized papain in cysteine/HCl-containing buffer at 37°C with gentle shaking. Digestion was monitored by taking aliquots and analyzing heavy chain cleavage using SDS-PAGE. To stop the reaction, immobilized papain was spun off and washed with 50 mM Tris pH 7.5 and filtered. Undigested full-length antibodies and Fc fragments were removed by using MabSelectSure (GE) columns.

The binding affinity of monoclonal antibodies and antibody fragments to HGF was assessed on OctetQK (ForteBio) using biolayer interferometry (BLI). Biotinylated antibodies or antibody fragments were immobilized (1 μg/mL for 500 seconds) onto the surface of streptavidin (SA) biosensors. A dilution series of human HGF (R&D Systems , Catalog #294-HGN/CF) to query antibodies. Various concentrations of antigen (ranging from about 80 ng/mL to 640 ng/mL) were used. Individual sensor data were fitted using a 1:1 Langmuir binding model under Octet analysis software (v3.1 ForteBio) using an association time of 15 minutes and a dissociation time of 35 minutes.

The kinetic binding values of Ab8, Ab10 and Ab12 to human-HGF determined as described above are included in the table below.

k _d (1/s) k _a (1/Ms) K _D (pM) Ab8 4.55E-05 3.73E+05 150 Ab10 5.13E-05 2.05E+05 250 Ab12 3.73E-05 2.76E+05 140

Example 10: Anti-tumor activity of recombinantly expressed anti-HGF antibody of the present invention

In vivo evaluation

In order to study the anti-tumor efficacy of anti-HGF antibody and the ability of anti-HGF antibody, a human glioma xenograft model implanted subcutaneously with U-87MG was used. Five-week-old athymic NCr/nu/nu mice were acclimatized for 16 days and subsequently inoculated by subcutaneous injection of U-87MG human glioma cells (ATCC, HTB-14, Lot 1653122) from in vitro cell culture. Each mouse received 20 million (2 x 107) U-87MG glioma cells resuspended in 0.2 mL MEM Eagle's medium (Eaglesmedia). Cell counts and viability were determined with a Beckman Coulter VICELL XR cell counter and viability analyzer. The day of tumor implantation was denoted as day 0. Tumor weights were brought to 162-294 mg (162-294 mm ³ in size) before treatment was initiated. Sufficient numbers of mice were implanted to select tumors with as narrow a weight change as possible for testing on the day of treatment initiation (day 18 after tumor implantation, expressed as SD). Those animals with an appropriate change in tumor size were selected and divided into treatment groups so that the median tumor weights were as close as possible to each other on the first day of treatment.

medical treatement

Regarding the antitumor and survival data corresponding to Figures 28 and 29, each group consisted of ten animals. All test compounds were administered by intraperitoneal (ip) injection at a dose of 10 mg/kg/injection twice every four days for five weeks (Q4D x 2/5 weeks).

For the antitumor and survival data corresponding to Figures 30 and 31; and 32 and 33, respectively, each group again consisted of ten animals. All test compounds were administered by intraperitoneal (ip) injection at a dose of 10 mg/kg/injection twice every four days for five weeks (Q4D x 2/5 weeks).

Regarding the antitumor and survival data corresponding to Figures 34 and 35, each group again consisted of ten animals. All test compounds were administered by intraperitoneal (ip) injection twice every four days for five weeks (Q4D x 2/5 weeks). The doses of Ab8 and Ab10 were 30, 10 or 2.5 mg/kg/injection. The dose of negative control antibody was 30 mg/kg/injection.

Tumor measurements and body weight

Subcutaneous tumors were measured and animals were weighed twice a week on the day of initiation of the first treatment. Tumor volumes were determined by caliper measurements (mm) and using the ellipsoid formula: L x W2/2 = ^mm3 , where L and W refer to the larger and smaller vertical dimensions collected under each measurement. This formula was also used to calculate tumor weight, assuming unit density (1 ^mm3 = 1 mg).

study duration

With respect to the data corresponding to Figures 28 and 29, the in vivo study was terminated 46 days after tumor implantation. Any animal found to be moribund or whose tumor reached 4,000 mg, ulcerated or discarded was euthanized prior to study termination. Figure 28 provides the response of subcutaneous U-87MG gliomas obtained after the protocol to treatment with negative control antibodies, Ab 10 or Ab 12 (10 mg/kg/injection). Figure 29 provides the survival ratio curves of subcutaneous U-87MG glioma treated with negative control antibody or Ab 10 or Ab 12 (10 mg/kg/injection) obtained after the protocol.

Regarding the antitumor and survival data corresponding to Figures 30 and 31 and 32 and 33, the in vivo study was terminated 79 days after tumor implantation. Any animal found to be moribund or whose tumor reached 4,000 mg, ulcerated or discarded was euthanized prior to study termination. Figure 30 provides subcutaneous U-87MG glioma responses to treatment with increasing doses of Ab8 (10, 2.5 and 0.25 mg/kg/injection) and negative control antibody (10 mg/kg/injection). Figure 31 presents a survival ratio curve for subcutaneous U-87MG glioma treated with increasing doses of Ab8 (10, 2.5 and 0.25 mg/kg/injection) or negative control antibody (10 mg/kg/injection).

Figure 32 provides subcutaneous U-87MG glioma responses to treatment with increasing doses of AblO (10, 2.5 and 0.25 mg/kg/injection) or negative control antibody (10 mg/kg/injection). Figure 33 provides a curve of the survival ratio of subcutaneous U-87MG glioma treated with increasing doses of AblO (10, 2.5 and 0.25 mg/kg/injection) or negative control antibody (10 mg/kg/injection).

With respect to the data corresponding to Figures 34 and 35, the in vivo study was terminated 80 days after tumor implantation. Any animal found to be moribund or whose tumor reached 4,000 mg, ulcerated or discarded was euthanized prior to study termination. Figure 34 provides the response of subcutaneous U-87MG gliomas obtained after the previous protocol to treatment with increasing doses of Ab28 (30, 10 and 2.5 mg/kg/injection) or negative control antibody (30 mg/kg/injection). Figure 35 provides the survival ratio of subcutaneous U-87MG glioma treated with increasing doses of Ab28 (30, 10 and 2.5 mg/kg/injection) or negative control antibody (30 mg/kg/injection) obtained after the previous regimen curve.

Example 11: Inhibition of c-met phosphorylation by recombinant expression of anti-HGF antibody of the present invention

The c-met receptor has several phosphorylation sites that have unique functions. Y1003 is in the juxtamembrane domain and complements the c-Cbl protein, which is involved in the ubiquitination of the receptor. It is said to be a negative regulatory site. Y1234/35 is the main site. Required for kinase activity and biological functions such as motility and morphogenesis. Y1349 and Y1356 serve as docking sites for protein-like PI3K and PLC-γ.

To investigate the ability of Ab8 to inhibit human HGF to direct Met phosphorylation, anti-phospho-Met western blots were performed as follows. Prostate cancer previously maintained in growth medium (F-12K, #30-2004, ATCC Manassas, VA.) containing 10% FBS (Hyclone, Victoria, Australia) was isolated using 0.25% trypsin (Hyclone, Logan, Utah). Confluent cultures of PC-3 cells (CRL-1435, ATCC, Manassas, VA.) were seeded at a density of 500,000 cells/well in 6-well plates. After overnight incubation at 37°C, the growth medium was removed and cells were serum starved overnight in FBS-free medium. By incubating 100 nM antibody and 1.25 nM HGF (GibcoLifeSciences, cat# PH60254) in serum-free medium in a 15 ml conical tube for 1 hour at 37°C, then added to the cells and incubating for an additional 10 minutes at 37°C, The medium was removed, rinsed with cold PBS and treated with supplemented protease inhibitor cocktail (Protease Inhibitors, Cat. No. 1836170, Roche, Indianapolis, IN. plus 10 mM NaF), Phosphatase Inhibitor Cocktail 1 (Cat. No. P-2850, Sigma , St.Louis, MO.) and Tris lysis buffer (catalog number R6OTX-2, MesoScaleDiscovery, Gaithersburg, MD) of phosphatase inhibitor cocktail 2 (catalog number P-5726, Sigma, St.Louis, MO.) Dissolved and assayed for inhibition of human HGF-dependent Met phosphorylation by Ab8 or a negative control antibody. Cell lysates were scraped from the 6-well plate and passed 5 times through a 23g needle while incubating on ice. Approximately 20 μg of total protein was loaded onto 4-12% Bis-trisNupage gels (Invitrogen, Carlsbad, CA) and subsequently transferred via iblot (Invitrogen, Carlsbad, CA). The nitrocellulose membrane was blocked with 3% BSA in TBS for 1 hour at room temperature with gentle shaking and the following antibodies (Cell Signaling Technologies, Beverly, Incubate overnight with one of: Phosphorylated Met(Y1234/35) Rabbit mAb (Cat. No. 3129, Cell Signaling Technologies, Beverly, Mass), Phosphorylated Met(Y1349) Rabbit mAb (Cat. No. 3133, Cell Signaling Technologies, Beverly, Mass) , GAPDHXP rabbit mAb (Catalog #5174, Cell Signaling Technologies, Beverly, Mass) or affinity purified rabbit anti-phospho-HGFR/c-met (Y1003) antibody (Cat #AF4059, R&D Systems, Minneapolis, MN). The nitrocellulose membrane was then washed 3 times in PBS + 0.05% Tween-20 and incubated at room temperature with a 1:10,000 dilution of peroxidase-conjugated affinity-purified goat anti-rabbit-specific IgG, Fc fragment (#111-035 -046, Jackson Immunoresearch, West Grove, PA) in 3% BSA, TBS+0.1% Tween-20+0.01% SDS for 2 hours. Then rinse 3 times as above. Finally, the cell membrane was developed for 5 minutes using SuperSignalWestPico chemiluminescent substrate and exposed to X-ray membrane. The results in Figure 36 show inhibition of human HGF driven phosphorylation of c-met by Ab8 using PC-3 cells (prostate cancer) Y1234/35, Y1003 and Y1349. The data herein shows that Ab8 exhibits inhibition of human HGF-dependent Met phosphorylation at all sites, whereas negative controls do not alter c-met phosphorylation.

Example 12: Effects of recombinantly expressed anti-HGF antibody of the present invention on cell proliferation

Proliferation assay

The effect of anti-HGF antibodies according to the invention on cell proliferation in vitro was analyzed. In these experiments, 4mBr-5 cells (rhesus bronchial epithelial cell line) were obtained from ATCC and used to characterize inhibition of HGF-driven cell proliferation by various antibody formulations. 4mBr-5 cells are a factor-dependent cell line that proliferates upon exposure to epidermal growth factor (EGF) or HGF.

4mBr-5 cells were grown in Ham's F-12K medium supplemented with 10% FBS and 50 ng/ml recombinant human EGF (GibcoLife Technologies). The cells were treated with 0.25% trypsin, washed twice with PBS and resuspended in Ham's F-12K medium supplemented with 2.5% FBS (assay medium). Cell density was determined using an Invitrogen Countess automated cell counter. Using 100 μl per well, cells were seeded at 200,000 cells/ml into clear bottom black walled 96 well plates (Costar) and incubated overnight. Assay medium supplemented with 100 ng/ml HGF (Gibco Life Sciences, catalog # PH60254) was incubated in the presence of 10 μg/ml negative control antibody or incubated for 1 hour at 37°C. Medium was removed and replaced with assay medium alone, which was then supplemented with 100 ng/ml HGF or 100 ng/ml HGF incubated in the presence of various antibodies. All conditions were performed in triplicate. Cells were allowed to proliferate for 48 hours. Media was removed and cells were washed twice with PBS. Cells were incubated with 4 μg/ml Calcein AM (Invitrogen) for 30 min at 37°C and fluorescence reading was performed on a Molecular Devices SpectraMax M2 using excitation/emission maxima 490/520 nm.

Figures 37-50 contain the results of these experiments, respectively. As shown herein, different anti-HGF antibodies (Abl, Ab2, Ab7, Ab8, Ab9, Ab10, Ab12, Ab14, Ab19, Ab21, Ab23, Ab24, Ab25 and Ab28) according to the invention inhibit the proliferation of 4mBr-5 cells .

Example 13: Inhibition of cell invasion by recombinant expression of anti-HGF antibody of the present invention

Cell Invasion Assay

The effect of anti-HGF antibodies according to the invention on cell invasion was tested. In these experiments, DBTRG cells (human glioblastoma cell line) were obtained from ATCC and cultured in RPMI-160 supplemented with 10% FBS, L-glutamine (Hyclone) and non-essential amino acids (Hyclone). Growth factor reduced Matrigel invasion chambers (24-well plate, BD Biosciences) were hydrated with 0.75 ml serum-free RPMI-1640 in the lower chamber and 0.5 ml serum-free RPMI-1640 in the insert. Incubate the chamber at 37 °C for 2 h. Media was removed from both the chamber and 0.75 ml serum-free RPMI1640 supplemented with 0.1% BSA and 10 ng/ml HGF or preincubated with 2 μg/ml negative control antibody or Ab8 at 37°C for 30 minutes. 10ng/mlHGF. All conditions were performed in duplicate. DBTRG cells were removed from PBS-supplemented plates using EDTA, pelleted by centrifugation and washed twice with PBS. Cells were then added to the insert chamber at a cell density of 20,000 cells/ml using 0.5 ml of serum-free medium. Incubate the chamber at 37°C for 24 hours. After incubation, medium was removed from the inserts and non-invasive cells were removed with a cotton swab. Inserts were fixed and stained using the Diff-Quick staining kit (source). Cell membranes were removed from the inserts, air-dried and invading cells counted by microscopy. As shown in Figure 39, Ab8 completely blocked DBTRG Matrigel invasion while the negative control antibody had no effect on cell invasion.

in conclusion

Described herein are novel anti-HGF antibodies and antibody fragments, nucleic acids, compositions thereof, and methods of use thereof, particularly for the treatment of indications when used as monotherapy or in combination with other therapies or agents. While the embodiments disclosed herein are preferred, those skilled in the art will appreciate that various substitutions, modifications, changes or changes may be made herein by those skilled in the art and are intended to be covered by the following claims.

The above description of various illustrated embodiments of the invention is not intended to be exhaustive or to limit the invention to the precise forms disclosed. While specific embodiments of, and examples for, the invention are described herein for illustrative purposes, various equivalent modifications are possible within the scope of the invention, as those skilled in the relevant art will recognize. The teachings of the invention provided herein may be adapted for purposes other than the embodiments described above.

These and other changes can be made to the invention in light of the above detailed description. Generally, in the following claims, the terms used should not be construed to limit the invention to the specific embodiments disclosed in the specification and the claims. Accordingly, the invention is not limited by the disclosure, but rather the scope of the invention is to be determined entirely by the following claims.

The invention may be practiced in other ways than those specifically described in the foregoing description and examples. Many modifications and variations of the present invention are possible in light of the above teaching and, therefore, are within the scope of the appended claims.

Certain teachings related to methods of obtaining a clonal population of antigen-specific B cells are disclosed in U.S. Provisional Patent Application No. 60/801,412, filed May 19, 2006, the disclosure of which is incorporated by reference in its entirety. into this article.

Certain teachings related to the humanization and preferred sequence modification of rabbit-derived monoclonal antibodies to maintain antigen binding affinity are disclosed in International Application No. PCT/US2008, filed May 21, 2008 entitled "Novel Rabbit Antibody Humanization Methods and Humanized Rabbit Antibodies" /064421 corresponds to International Publication No. WO/2008/144757, the disclosure of which is incorporated herein by reference in its entirety.

Certain teachings related to the production of antibodies or fragments thereof using competent mating yeast and corresponding methods are disclosed in U.S. Patent Application No. 11/429,053, filed May 8, 2006 (U.S. Patent Application Publication No. US2006/0270045 No.), the disclosure of which is incorporated herein by reference in its entirety.

Certain HGF antibody polynucleotides and polypeptides are disclosed in the Sequence Listing accompanying this patent application file, and the disclosure of said Sequence Listing is incorporated herein by reference in its entirety.

The entire disclosures of each of the documents (including patents, patent applications, journal articles, abstracts, manuals, books or other disclosures) cited in the Background of the Invention, Detailed Description and Examples are incorporated herein by reference in their entirety.

Claims (23)

1. anti-human pHGF (HGF) antibody or an antibody fragment, its:

(1) epi-position identical or overlapping with the anti-human HGF antibody of Ab28 with being selected from Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and/or competition binding extremely described identical or overlapping epi-position on specific binding to whole person HGF polypeptide or its fragment;

(2) identical with the anti-human HGF antibody of Ab28 with being selected from Ab1, Ab2, Ab7, Ab8, Ab9, Ab10, Ab12, Ab13, Ab14, Ab19, Ab21, Ab23, Ab24, Ab25 on specific binding to whole person HGF polypeptide or its fragment linear or conformational epitope and/or competition binding are to the epi-position identical or overlapping with described antibody;

(3) identical with the anti-human HGF antibody of Ab28 with being selected from Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 on specific binding to whole person HGF polypeptide or its fragment epi-position; And/or

(4) on specific binding to whole person HGF polypeptide or its fragment be selected from Ab1, Ab2, Ab7, Ab8, Ab9, Ab10, Ab12, Ab13, Ab14, Ab19, Ab21, Ab23, Ab24, the epi-position that the anti-human HGF antibody of Ab25 with Ab28 is identical, wherein said epi-position optionally uses to combine and measures and differentiated, described combination measures the combination of one or more peptides in the library detecting described anti-human HGF antibody and 10-15-mer peptide, described 10-15-mer peptide is correspond to overlapping peptide fragments that is all or the people HGF of all described people HGF polypeptide length substantially, wherein combine and measure further optionally for protein immunoblotting measures, its by use chemiluminescent labeling to detect in described antibody or antibody fragment and described library one or more described in the specific binding of 10-15mer peptide, detectable chemiluminescence signal is launched during the peptide of described chemiluminescent labeling in described antibody or antibody fragment specific binding to described library.

2. anti-human HGF antibody as claimed in claim 1 or antibody fragment, wherein:

(1) described anti-human HGF antibody or antibody fragment contain at least 2 complementary determining regions (CDR) be selected from by the anti-HGF antibody in following formed group: Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28;

(2) described anti-human HGF antibody or antibody fragment contain at least 3 complementary determining regions (CDR) be selected from by the anti-HGF antibody in following formed group: Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28;

(3) described anti-human HGF antibody or antibody fragment contain at least 4 complementary determining regions (CDR) be selected from by the anti-HGF antibody in following formed group: Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28;

(4) described anti-human HGF antibody or antibody fragment contain at least 5 complementary determining regions (CDR) be selected from by the anti-HGF antibody in following formed group: Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28; Or

(5) described anti-human HGF antibody or antibody fragment contain whole 6 complementary determining regions (CDR) be selected from by the anti-HGF antibody in following formed group: Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28;

3. anti-human HGF antibody as claimed in claim 1 or 2 or antibody fragment, wherein:

(1) described anti-human HGF antibody contains at least one residue except CDR residue, and at least one residue described is modified, and described modification does not affect HGF and combines;

(2) described anti-human HGF antibody is chimeric and contains people's constant domain;

(3) described anti-HGF antibody is behaved or humanized antibody or its fragment;

(4) described anti-human HGF antibody or antibody fragment contain:

(a) variable heavy chain, it comprises and is selected from by SEQIDNO:4, SEQIDNO:44, SEQIDNO:84, SEQIDNO:124, SEQIDNO:164, SEQIDNO:204, SEQIDNO:244, SEQIDNO:284, SEQIDNO:324, SEQIDNO:364, SEQIDNO:404, SEQIDNO:444, SEQIDNO:484, SEQIDNO:524, SEQIDNO:564, SEQIDNO:604, SEQIDNO:644, SEQIDNO:684, SEQIDNO:724, SEQIDNO:764, SEQIDNO:804, SEQIDNO:844, SEQIDNO:884, SEQIDNO:924, SEQIDNO:964, the CDR1 sequence of the group of SEQIDNO:1004 and SEQIDNO:1044 composition, be selected from by SEQIDNO:6, SEQIDNO:46, SEQIDNO:86, SEQIDNO:126, SEQIDNO:166, SEQIDNO:206, SEQIDNO:246, SEQIDNO:286, SEQIDNO:326, SEQIDNO:366, SEQIDNO:406, SEQIDNO:446, SEQIDNO:486, SEQIDNO:526, SEQIDNO:566, SEQIDNO:606, SEQIDNO:646, SEQIDNO:686, SEQIDNO:726, SEQIDNO:766, SEQIDNO:806, SEQIDNO:846, SEQIDNO:886, SEQIDNO:926, SEQIDNO:966, the CDR2 sequence of the group of SEQIDNO:1006 and SEQIDNO:1046 composition, with be selected from by SEQIDNO:8, SEQIDNO:48, SEQIDNO:88, SEQIDNO:128, SEQIDNO:168, SEQIDNO:208, SEQIDNO:248, SEQIDNO:288, SEQIDNO:328, SEQIDNO:368, SEQIDNO:408, SEQIDNO:448, SEQIDNO:488, SEQIDNO:528, SEQIDNO:568, SEQIDNO:608, SEQIDNO:648, SEQIDNO:688, SEQIDNO:728, SEQIDNO:768, SEQIDNO:808, SEQIDNO:848, SEQIDNO:888 and SEQIDNO:928, SEQIDNO:968, the CDR3 sequence of the group of SEQIDNO:1008 and SEQIDNO:1048 composition, and/or

(b) variable light, it comprises and is selected from by SEQIDNO:24, SEQIDNO:64, SEQIDNO:104, SEQIDNO:144, SEQIDNO:184, SEQIDNO:224, SEQIDNO:264, SEQIDNO:304, SEQIDNO:344, SEQIDNO:384, SEQIDNO:424, SEQIDNO:464, SEQIDNO:504, SEQIDNO:544, SEQIDNO:584, SEQIDNO:624, SEQIDNO:664, SEQIDNO:704, SEQIDNO:744, SEQIDNO:784, SEQIDNO:824, SEQIDNO:864, SEQIDNO:904, SEQIDNO:944, SEQIDNO:984, the CDR1 sequence of the group of SEQIDNO:1024 and SEQIDNO:1064 composition, be selected from by SEQIDNO:26, SEQIDNO:66, SEQIDNO:106, SEQIDNO:146, SEQIDNO:186, SEQIDNO:226, SEQIDNO:266, SEQIDNO:306, SEQIDNO:346, SEQIDNO:386, SEQIDNO:426, SEQIDNO:466, SEQIDNO:506, SEQIDNO:546, SEQIDNO:586, SEQIDNO:626, SEQIDNO:666, SEQIDNO:706, SEQIDNO:746, SEQIDNO:786, SEQIDNO:826, SEQIDNO:866, SEQIDNO:906, SEQIDNO:946, SEQIDNO:986, the CDR2 sequence of the group of SEQIDNO:1026 and SEQIDNO:1066 composition, with be selected from by SEQIDNO:28, SEQIDNO:68, SEQIDNO:108, SEQIDNO:148, SEQIDNO:188, SEQIDNO:228, SEQIDNO:268, SEQIDNO:308, SEQIDNO:348, SEQIDNO:388, SEQIDNO:428, SEQIDNO:468, SEQIDNO:508, SEQIDNO:548, SEQIDNO:588, SEQIDNO:628, SEQIDNO:668, SEQIDNO:708, SEQIDNO:748, SEQIDNO:788, SEQIDNO:828, SEQIDNO:868, SEQIDNO:908, SEQIDNO:948, SEQIDNO:988, the CDR3 sequence of the group of SEQIDNO:1028 and SEQIDNO:1068 composition, one or two residue that its further restricted condition is arbitrary previously differentiated CDR polypeptide through another aminoacid replacement, can be preferably conservative amino acid and replaces,

(5) the described variable heavy chain of described antibody or antibody fragment comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:4, the CDR2 sequence of SEQIDNO:6 and SEQIDNO:8; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:24, the CDR2 sequence of SEQIDNO:26 and SEQIDNO:28;

(6) described anti-human HGF antibody or antibody fragment contain variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:44, the CDR2 sequence of SEQIDNO:46 and SEQIDNO:48; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:64, the CDR2 sequence of SEQIDNO:66 and SEQIDNO:68;

(7) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:84, the CDR2 sequence of SEQIDNO:86 and SEQIDNO:88; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:104, the CDR2 sequence of SEQIDNO:106 and SEQIDNO:108;

(8) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:124, the CDR2 sequence of SEQIDNO:126 and SEQIDNO:128; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:144, the CDR2 sequence of SEQIDNO:146 and SEQIDNO:148;

(9) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:164, the CDR2 sequence of SEQIDNO:166 and SEQIDNO:168; And described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:184, the CDR2 sequence of SEQIDNO:186 and SEQIDNO:188;

(10) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:204, the CDR2 sequence of SEQIDNO:206 and SEQIDNO:208; And described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:224, the CDR2 sequence of SEQIDNO:226 and SEQIDNO:228;

(11) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:244, the CDR2 sequence of SEQIDNO:246 and SEQIDNO:248; And described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:264, the CDR2 sequence of SEQIDNO:266 and SEQIDNO:268;

(12) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:284, the CDR2 sequence of SEQIDNO:286 and SEQIDNO:288; And described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:304, the CDR2 sequence of SEQIDNO:306 and SEQIDNO:308;

(13) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:324, the CDR2 sequence of SEQIDNO:326 and SEQIDNO:328; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:344, the CDR2 sequence of SEQIDNO:346 and SEQIDNO:348;

(14) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:364, the CDR2 sequence of SEQIDNO:366 and SEQIDNO:368; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:384, the CDR2 sequence of SEQIDNO:386 and SEQIDNO:388;

(15) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:404, the CDR2 sequence of SEQIDNO:406 and SEQIDNO:408; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:424, the CDR2 sequence of SEQIDNO:426 and SEQIDNO:428;

(16) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:444, the CDR2 sequence of SEQIDNO:446 and SEQIDNO:448; And described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:464, the CDR2 sequence of SEQIDNO:466 and SEQIDNO:468;

(17) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:484, the CDR2 sequence of SEQIDNO:486 and SEQIDNO:488; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:504, the CDR2 sequence of SEQIDNO:506 and SEQIDNO:508;

(18) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:524, the CDR2 sequence of SEQIDNO:526 and SEQIDNO:528; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:544, the CDR2 sequence of SEQIDNO:546 and SEQIDNO:548;

(19) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:564, the CDR2 sequence of SEQIDNO:566 and SEQIDNO:568; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:584, the CDR2 sequence of SEQIDNO:586 and SEQIDNO:588;

(20) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:604, the CDR2 sequence of SEQIDNO:606 and SEQIDNO:608; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:624, the CDR2 sequence of SEQIDNO:626 and SEQIDNO:628;

(21) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:644, the CDR2 sequence of SEQIDNO:646 and SEQIDNO:648; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:664, the CDR2 sequence of SEQIDNO:666 and SEQIDNO:668;

(22) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:684, the CDR2 sequence of SEQIDNO:686 and SEQIDNO:688; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:704, the CDR2 sequence of SEQIDNO:706 and SEQIDNO:708;

(23) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:724, the CDR2 sequence of SEQIDNO:726 and SEQIDNO:728; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:744, the CDR2 sequence of SEQIDNO:746 and SEQIDNO:748;

(24) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, it comprises anti-human HGF antibody according to any one of claim 1 to 10 or antibody fragment, and wherein said variable heavy chain comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:764, the CDR2 sequence of SEQIDNO:766 and SEQIDNO:768; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:784, the CDR2 sequence of SEQIDNO:786 and SEQIDNO:788;

(25) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:804, the CDR2 sequence of SEQIDNO:806 and SEQIDNO:808; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:824, the CDR2 sequence of SEQIDNO:826 and SEQIDNO:828;

(26) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:844, the CDR2 sequence of SEQIDNO:846 and SEQIDNO:848; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:864, the CDR2 sequence of SEQIDNO:866 and SEQIDNO:868;

(27) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:884, the CDR2 sequence of SEQIDNO:886 and SEQIDNO:888; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:904, the CDR2 sequence of SEQIDNO:906 and SEQIDNO:908;

(28) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:924, the CDR2 sequence of SEQIDNO:926 and SEQIDNO:928; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:944, the CDR2 sequence of SEQIDNO:946 and SEQIDNO:948;

(29) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:964, the CDR2 sequence of SEQIDNO:966 and SEQIDNO:968; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:984, the CDR2 sequence of SEQIDNO:986 and SEQIDNO:988;

(30) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:1004, the CDR2 sequence of SEQIDNO:1006 and SEQIDNO:1008; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:1024, the CDR2 sequence of SEQIDNO:1026 and SEQIDNO:1028;

(31) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:1044, the CDR2 sequence of SEQIDNO:1046 and SEQIDNO:1048; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:1064, the CDR2 sequence of SEQIDNO:1066 and SEQIDNO:1068;

(32) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:2 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and/or described variable light comprise SEQIDNO:22;

(33) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:2 with SEQIDNO:22 respectively separately;

(34) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:42 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:62 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(35) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:42 with SEQIDNO:62 respectively separately;

(36) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:82 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:102 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(37) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:82 with SEQIDNO:102 respectively separately;

(38) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:122 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:142 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(39) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:122 with SEQIDNO:142 respectively separately;

(40) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:162 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:182 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(41) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:162 with SEQIDNO:182 respectively separately;

(42) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:202 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:222 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(43) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:202 with SEQIDNO:222 respectively separately;

(44) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:242 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:262 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(45) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:242 with SEQIDNO:262 respectively separately;

(46) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:282 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:302 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(47) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:282 with SEQIDNO:302 respectively separately;

(48) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:322 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:342 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(49) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:322 with SEQIDNO:342 respectively separately;

(50) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:362 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:382 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(51) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:362 with SEQIDNO:382 respectively separately;

(52) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:402 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:422 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(53) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:402 with SEQIDNO:422 respectively separately;

(54) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:442 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:462 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(55) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:442 with SEQIDNO:462 respectively separately;

(56) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:482 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:502 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(57) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:482 with SEQIDNO:502 respectively separately;

(58) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:522 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:542 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(59) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:522 with SEQIDNO:542 respectively separately;

(60) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:562 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:582 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(61) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:562 with SEQIDNO:582 respectively separately;

(62) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:602 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:622 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(63) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:602 with SEQIDNO:622 respectively separately;

(64) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:642 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:662 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(65) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:642 with SEQIDNO:662 respectively separately;

(66) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:682 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:702 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(67) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:682 with SEQIDNO:702 respectively separately;

(68) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:722 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:742 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(69) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:722 with SEQIDNO:742 respectively separately;

(70) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:762 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:782 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(71) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:762 with SEQIDNO:782 respectively separately;

(72) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:802 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:822 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(73) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:802 with SEQIDNO:822 respectively separately;

(74) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:842 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:862 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(75) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:842 with SEQIDNO:862 respectively separately;

(76) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:882 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:902 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(77) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:882 with SEQIDNO:902 respectively separately;

(78) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:922 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:942 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(79) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:922 with SEQIDNO:942 respectively separately;

(80) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:962 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:982 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(81) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:962 with SEQIDNO:982 respectively separately;

(82) described anti-human HGF antibody or antibody fragment variable heavy chain, its comprise with SEQIDNO:1002 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:1022 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(83) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:1002 with SEQIDNO:1022 respectively separately;

(84) described anti-human HGF antibody or antibody fragment comprise variable heavy chain, its comprise with SEQIDNO:1042 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described variable light comprise with SEQIDNO:1062 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(85) described anti-human HGF antibody or antibody fragment comprise variable heavy chain and light chain, and it is identical with light chain at least 90% with the variable heavy chain in SEQIDNO:1042 with SEQIDNO:1062 respectively separately;

(86) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:1 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprise with SEQIDNO:21 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(87) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:41 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence; And light chain, its comprise with SEQIDNO:61 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(88) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:81 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence; And light chain, its comprise with SEQIDNO:101 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(89) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:121 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence; And light chain, its comprise with SEQIDNO:141 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(90) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:161 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprise with SEQIDNO:181 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(91) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:201 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprise with SEQIDNO:221 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(92) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:241 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprise with SEQIDNO:261 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(93) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:281 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprises SEQIDNO:301;

(94) described anti-human HGF antibody or antibody fragment comprise heavy chain, and it comprises SEQIDNO:321; And light chain, its comprise with SEQIDNO:341 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(95) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:361 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprises SEQIDNO:381;

(96) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise SEQIDNO:401 and described light chain comprise with SEQIDNO:421 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(97) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:441 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprise with SEQIDNO:461 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(98) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:481 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprise with SEQIDNO:501 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(99) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:521 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprises SEQIDNO:541;

(100) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise SEQIDNO:561 and described light chain comprise with SEQIDNO:581 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(101) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:601 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprises SEQIDNO:621;

(102) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise SEQIDNO:641 and described light chain comprise with SEQIDNO:661 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(103) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:681 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprise with SEQIDNO:701 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(104) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:721 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprises SEQIDNO:741;

(105) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise SEQIDNO:761 and described light chain comprise with SEQIDNO:781 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(106) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:801 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprise with SEQIDNO:821 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(107) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:841 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprise with SEQIDNO:861 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(108) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise with SEQIDNO:881 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence and described light chain comprises SEQIDNO:901;

(109) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise SEQIDNO:921 and described light chain comprise with SEQIDNO:941 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(110) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise SEQIDNO:961 and described light chain comprise with SEQIDNO:981 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(111) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise SEQIDNO:1001 and described light chain comprise with SEQIDNO:1021 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence; With

(112) described anti-human HGF antibody or antibody fragment comprise heavy chain, its comprise SEQIDNO:1041 and described light chain comprise with SEQIDNO:1061 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence; Or any combination of above-mentioned each.

4. the anti-human HGF antibody as described in claim 1,2 or 3 or antibody fragment, wherein said antibody or antibody fragment comprise:

(1) chimeric, humanization and people's antibody or antibody fragment;

(2) scFv, camel antibodies, nano antibody, IgNAR, Fab fragment, Fab' fragment, MetMab sample antibody, monovalent antibody fragments and/or F (ab') 2 fragment;

(3) N-glycosylation and/or the glycosylated antibody of O-or antibody fragment is lacked substantially or completely;

(4) antibody or the antibody fragment of people's constant domain is comprised;

(5) people containing human IgG1, IgG2, IgG3 or IgG4 antibody constant region;

(6) antibody or the antibody fragment in Fc district is comprised, described Fc district at least one to change in effector function, transformation period, proteolysis or glycosylation modified;

(7) antibody containing Fc district, described Fc district contains change or eliminates N-glycosylation and/or the glycosylated one or more sudden change of O-;

(8) humanized antibody or antibody fragment;

(9) to be less than or equal to 10 ^-2m, 5 × 10 ^-3m, 10 ^-3m, 5 × 10 ^-4m, 10 ^-4m, 5 × 10 ^-5m, 10 ^-5m, 5 × 10 ^-6m, 10 ^-6m, 5 × 10 ^-7m, 10 ^-7m, 5 × 10 ^-8m, 10 ^-8m, 5 × 10 ^-9m, 10 ^-9m, 5 × 10 ^-10m, 10 ^-10m, 5 × 10 ^-11m, 10 ^-11m, 5 × 10 ^-12m, 10 ^-12m, 5 × 10 ^-13m or 10 ^-13dissociation constant (the K of M _d) be bonded to antibody or the antibody fragment of HGF;

(10) to be less than or equal to 10 ^-11m, 5 × 10 ^-12m or 10 ^-12dissociation constant (the K of M _d) be bonded to antibody or the antibody fragment of HGF;

(11) to be less than or equal to 10 ^-4s ^-1, 5 × 10 ^-5s ^-1, 10-5S ^-1, 5 × 10 ^-6s ^-1, 10 ^-6s-1,5 × 10 ^-7s ^-1or 10-7S ^-1dissociation rate (K _off) be bonded to antibody or the antibody fragment of HGF;

(12) antibody or the antibody fragment of detectable label or therapeutical agent is directly or indirectly connected to;

(13) suppress when being applied to people experimenter or in and the antibody of at least one biological effect to be caused by HGF or antibody fragment;

(14) to be less than the K of about 100nM _dbe bonded to antibody or the antibody fragment of HGF;

(15) to be less than the K of about 10nM _dbe bonded to antibody or the antibody fragment of HGF;

(16) to be less than the K of about 1nM _dbe bonded to antibody or the antibody fragment of HGF;

(17) with between about 1 and K about between 10nM _dbe bonded to antibody or the antibody fragment of HGF;

(18) with between about 0.1 and K about between 1nM _dbe bonded to antibody or the antibody fragment of HGF;

(19) be connected at least one effect part, optionally comprise antibody or the antibody fragment of chemical linker;

(20) be connected to one or more can the antibody of test section or antibody fragment;

(21) being connected to can the antibody of test section or antibody fragment, describedly can comprise fluorescence dye, enzyme, substrate, noclilucence material, radioactive substance, chemiluminescent moiety or its mixture in test section;

(22) antibody or the antibody fragment of one or more funtion part is connected to; And/or

(23) for the antiidiotypic antibody produced according to anti-human HGF antibody or the antibody fragment of aforementioned any one.

5. be applicable to treat, prevention or the composition of diagnostic uses, it comprises treatment, prevention or the antibody diagnosing at least one of significant quantity according to any one of Claims 1-4 or antibody fragment.

6. composition as claimed in claim 5, wherein:

(1) it is applicable to by being selected from following mode to use: in cheek, epidermis, epidural, suction, intra-arterial, heart, in Intraventricular, intracutaneous, intramuscular, nose, in intraocular, intraperitoneal, backbone, in sheath, intravenously, per os, parenteral, locally, via under enema or suppository per rectum, subcutaneous, corium, sublingual, through skin with through mucous membrane;

(2) it comprises pulvis, liquid, gel, drops, liposome or other formulation;

(3) it is through freeze-drying;

(4) it comprises pharmaceutically acceptable thinner, carrier, solubilizing agent, emulsifying agent, sanitas or its mixture;

(5) it comprises another promoting agent;

(6) it comprises another promoting agent, and another promoting agent described comprises the medicine being applicable to treat the symptom being selected from cancer, macular degeneration, Alzheimer and malaria infection;

(7) it comprises another promoting agent being used for the treatment of and being selected from following cancer: the brain tumor (such as glioblastoma multiforme) of ovarian cancer, mammary cancer, lung cancer (minicell or non-small cell), colon and colorectal carcinoma, prostate cancer, carcinoma of the pancreas, kidney, cancer of the stomach, liver cancer, bladder cancer, thyroid carcinoma, carcinoma of endometrium, tumor of head and neck, melanoma, sarcoma, leukemia, lymphoma and children or adult;

(8) it comprises and is selected from another following promoting agent: chemotherapeutic, Statins, cytokine, immunosuppressor, gene therapeutic agents, anti-coagulant, anti-emaciation agent, anti-unable dose, fatigue protective agent, anti-heat-generating agent, nausea agent, antiemetic, IL-6 antagonist, cytotoxic agent, analgesic agent, febrifuge, antiphlogistic, microbiotic, antiviral agent, antibacterial agent agent, anti-angiogenic agent or its any combination;

(9) it comprises another promoting agent, another promoting agent described is be selected from following chemotherapeutic: VEGF antagonist, EGFR antagonist, platinum class, PTX class, irinotecan, 5 FU 5 fluorouracil, gemcitabine, formyl tetrahydrofolic acid, steroid, endoxan, melphalan, vinca alkaloids, vinealeucoblastine(VLB), vincristine(VCR), vindesine, vinorelbine, nitrogen mustards, tyrosine kinase inhibitor, radiotherapy dose, sexual hormoue antagonist, SARM, selective estrogen receptor modulators, PDGF antagonist, TNF antagonist, IL-I antagonist, interleukin, IL-12, IL-2, IL-12R antagonist, the monoclonal antibody that toxin is puted together, specific for tumour antigen monoclonal antibody, Erbitux ^tM, Avastin ^tM, handkerchief trastuzumab, anti-CD20 antibodies, auspicious pearl monoclonal antibody difficult to understand, method difficult to understand wood monoclonal antibody, DXL625, or its any combination,

(10) it comprises another promoting agent, and another promoting agent described is be selected from following anti-coagulant: ReoPro (ReoPro ^tM), acenocoumarol, antithrombin IE, argatroban, acetylsalicylic acid, Bivalirudin (Angiomax ^tM), clopidogrel, dabigatran, dabigatran etcxilate (Pradaxa ^tM/ Pradax ^tM), desirudin (Revasc ^tM/ Iprivask ^tM), Dipyridamole, eptifibatide (Integrilin ^tM), fondaparin, heparin, r-hirudin, according to DALT, Lepirudin (Refludan ^tM), low molecular weight heparin, Melagatran, phenindione, phenprocoumon, ticlopidine, Tirofiban (Aggrastat ^tM), warfarin, ximelagatran, ximelagatran (Exanta ^tM/ Exarta ^tM) or its any combination;

(11) it comprises another promoting agent, and another promoting agent described is be selected from following statin: atorvastatin, Cerivastatin, fluvastatin, lovastatin, mevastatin, Pravastatin, Pravastatin, Rosuvastatin, Simvastatin or its any combination;

(12) it comprises another promoting agent, and another promoting agent described is the antagonist being selected from the following factor: tumor necrosis factor-alpha, interferon-gamma, interleukin-11 α, interleukin-11 β, interleukin 6, proteolysis inducible factor, leukaemia inhibitory factor or its any combination;

(13) it comprises another promoting agent, another promoting agent described is be selected from following anti-angiogenesis: soluble VEGFR-1, NRP-1, ANG2, TSP-1 and TSP-2, angiostatin and associated molecule, endostatin, vascularization statin, calprotectin, PF4, TIMP, CDAI, Meth-1, Meth-2, IFN-α, IFN-β and IFN-γ, CXCL10, IL-4, IL-12, IL-18, thrombogen (kringle structural domain-2), Antithrombin III fragment, prolactin antagonist, VEGI, SPARC, osteopontin, mammary gland serine protease inhibitor and canstatin,

(14) it comprises another promoting agent, and another promoting agent described is comprise following anti-emaciation agent: hemp, dronabinol (Marinol ^tM), nabilone (Cesamet), cannabidiol, cannabichromene, tetrahydrocannabinol, Sativex, megestrol acetate or its any combination;

(15) it comprises another promoting agent, another promoting agent described is nausea agent or antiemetic, and it comprises: 5-HT3 receptor antagonist, asafoetide, alizapride, anticholinergic, antihistamine, Aprepitant, benzodiazepine class, cannabichromene, cannabidiol, cannaboid, hemp, Carcel smooth, chlorpromazine, marezine, dexamethasone, dexamethasone, umine (Gravol ^tM), diphenhydramine, dolasetron, domperidone, dopamine antagonist, doxylamine, dronabinol (Marinol ^tM), droperidol, more tell peaceful, ginger, granisetron, haloperidol, hydroxyzine, the sweet henbane alkali in east, lorazepam, Meclozine, Reglan, midazolam, muscimol, nabilone (Cesamet), nkl receptor antagonist, ondansetron, Palonosetron, Mentha arvensis L. syn.M.haplocalyxBrig, promethazine hydrochloride, prochlorperazine, Promacot, promethazine, Pentazine, Disoprofol, sativex, tetrahydrocannabinol, trimethobenzamide, tropisetron, nandrolone, stilboestrol, thalidomide, Revlimid, Leptin agonist, muscle mass antagonist, anti-muscle mass antibody, SARM, selective estrogen receptor modulators, Angiotensin All antagonist, beta 2 adrenoreceptor agonists, β 3 adrenoceptor agonists, or its any combination,

(16) it comprises another promoting agent, another promoting agent described is selected from tamoxifen, BCL-2 antagonist, oestrogenic hormon, diphosphonate, teriparatide, strontium ranelate, alendronate sodium (Fosamax), risedronate (safe and reliable good), raloxifene, ibandronate (Bang Luoli), Ao Bakela, ABT-263, gossypol, Gefitinib, epidermal growth factor recipient tyrosine kinase inhibitor, erlotinib, epidermal growth factor receptor inhibitor, psoralene, neosoralen, soloxsalen, bergapton, class xanthoplane, etretinate, Etretin, infliximab adalimumab, infliximab, etanercept, Zenapax ^tM, S-Neoral, methotrexate, G-CSF, filgrastim, lenograstim, excellent Bao Jin, Niu Lasita, 2-arylpropionic acid, Aceclofenac, acemetacin, acetylsalicylic acid (acetylsalicylic acid), Warner-Lambert), alminoprofen, amoxycilline Trihydrate bp, aminoantipyrene, aryl alkanoic acid, azapropazone, Benorilate/Benorilate, Compd 90459, Bromfenac, carprofen, celecoxib, choline salicylate magnesium, Perclusone, cox 2 inhibitor, Seractil, dexketoprofen, diclofenac, diflunisal, Droxicam, ethenzamide, R-ETODOLAC, Etoricoxib, Fes amine, that acid fragrant, fenbufen, fenoprofen, Flufenamic Acid, flunoxaprofen, flurbiprofen, Ibuprofen BP/EP, ibuproxam, indomethacin, indoprofen, recheton, Ketoprofen BP 93, ketorolac, lornoxicam, loxoprofen, lumiracoxib, magnesium salicylate, meclofenamic acid, mefenamic acid, meloxicam, Sulpyrine, wintergreen oil, mofebutazone, nabumetone, Naproxen Base, the adjacent amido phenylformic acid of N-aryl, oxametacin, Taisho), former times health class, Tacote, parecoxib, phenazone, phenylbutazone, phenylbutazone, piroxicam, pirprofen, profen, proglumetacin, pyrazolidine derivatives, rofecoxib, sasapyrin, salicylic amide, salicylate, sulfinpyrazone, sulindac, sutoprofen, tenoxicam, tiaprofenic acid, tolfenamic acid, tolmetin and valdecoxib, microbiotic, amikacin, aminoglycoside, amoxycilline Trihydrate bp, Ampicillin Trihydrate, ansamycins, Arsphenamine, Azythromycin, azlocillin, aztreonam, bacitracin, carbacephem, carbapenems, Pyocianil, cefaclor, S 578, Cephalexin Monohydrate Micro/Compacted, cefoxitin, cefoxitin, Cefamandole, Cephazolin, Cefdinir, cefditoren, cefepime, Cefixime Micronized, cefoperazone, cefotaxime, cefoxitin, Cefpodoxime, cefprozil, ceftazime, ceftibuten, ceftizoxime, Ceftobiprole, ceftriaxone, cephalofruxin, cephalosporin class, paraxin, cilastatin, Ciprofloxacin, clarithromycin, clindamycin, cloxacillin, Totazina, trimethoprim-sulfamethoxazole, dalfopristin, Demethylchlortetracycline, dicloxacillin, dirithromycin, doripenem, Vibravenos, enoxacin, ertapenem, erythromycin, Tibutol, Flucloxacillin, phosphonomycin, Nifurazolidone, fusidinic acid, Gatifloxacin, geldanamycin, gentamicin, glycopeptide class, herbimycin, imipenum, vazadrine, kantlex, levofloxacin, lincomycin, Linezolid, lomefloxacin, Loracarbef, Macrolide, mafenide, meropenem, X-1497, metronidazole, mezlocillin, MINOCYCLINE HCL, monobactam class, Moxifloxacin, mupirocin, nafcillin, Liu Suanyan NEOMYCIN SULPHATE, netilmicin, furadantin, norfloxicin, Ofloxacine USP 23, Oxazacillin, terramycin, paromycin, penicillin, penicillins, piperacillin, dull and stereotyped mycin, PXB, polypeptide, Prontosil, pyrazinoic acid amide, quinolones, Quinupristin, Rifampin, Rifampin, Roxithromycin, spectinomycin, Streptomycin sulphate, Sulphacetamide, ayerlucil, sulfanilamide (SN), sulfasalazine, Sulfafurazole, sulfamido, teicoplanin, Ketek, tsiklomitsin, tetracyclines, ticarcillin, fasigyne, tobramycin, Trimethoprim BP, Trimethoprim BP-sulfamethoxazole, troleomycin, trovafloxacin, vancomycin, aldosterone, beclometasone, Betamethasone Valerate, reflunomide, hydrocortisone, acetic acid cortisone, acetic acid Desoxycortone, dexamethasone, acetic acid fluohydrocortisone, glucocorticoids, hydrocortisone, methylprednisolone, prednisolone, prednisone, steroid and triamcinolone, antiviral agent, comprises such as Abacavir, acyclovir, acyclovir, Adefovir, amantadine, amprenavir, antiretroviral fixed dosage combines, antiretroviral works in coordination with toughener, Arbidol, Reyataz R, atorvastatincalcuim, brivudine, cidofovir, Combivir, DRV, Delavirdine, Didanosine, how can husky promise, edoxudine, efavirenz, emtricitabine, enfuirtide, Entecavir, entry inhibitor, Famciclovir, Fomivirsen, Fosamprenavir, FOSCARNET, PHOSPHONACETIC, fusion inhibitor, ganciclovir, Jia Dexi, ibacitabine, iodoxuridine, Imiquimod, isoprinosine, Indinavir, inosine, integrase inhibitor, Interferon, rabbit, I type Interferon, rabbit, II type Interferon, rabbit, type iii interferon, lamivudine, rltonavir, loviride, Maraviroc, MK-0518, Moroxydine, viracept see nelfinaivr, nevirapine, Nexavar, nucleoside analog, Oseltamivir, Penciclovir, Peramivir, Pu Kenali, podophyllotoxin, proteinase inhibitor, reverse transcriptase inhibitors, virazole, Rimantadine, ritonavir, Saquinavir, stavudine, tynofovir, tenofovir disoproxil, tipranavir, trifluridine, three associations only, tromantadine, Troyes reaches, valacyclovir, valganciclovir, Wei Kelinuo, vidarabine, the big miaow that draws is fixed, zalcitabine, zanamivir, zidovudine, or its any combination,

(17) it comprises another promoting agent, another promoting agent described is cytotoxic agent, chemotherapeutic or immunosuppressor, and it comprises: 1-boldenone, 1-methyl-nitroso-urea, 5 FU 5 fluorouracil, Ismipur, Ismipur, 6-Tioguanine, Orencia, abraxane, Etretin, aclarubicin, actinium-225 ( ²²⁵ac), actinomycin, adalimumab, adenosine deaminase inhibitors, Afelimomab, VEGF Trap, atropic pearl monoclonal antibody, Ah method's Saite, alitretinoin, alkyl sulfonate esters, alkylating agent, altretamine, ground, Avobenzene west, amino-laevulic acid/amino-laevulic acid methyl esters, aminopterin-induced syndrome, aminopterin-induced syndrome, amrubicin, amsacrine, amsacrine, anagrelide, Kineret, amerantrone class, anthracyclines, anthracyclines, anthracyclines, Antramycin (AMC), antimitotic agent, microbiotic, anti-CD20 antibodies, antifol, antilymphocyte globulin (ALG), antimetabolite, antithymocyte globulin, white arsenic, A Sai pearl monoclonal antibody, asparaginase, fall l-asparagine medicine, astatine-211 ( ²¹¹at), A Li pearl monoclonal antibody, atorolimumab, atrasentan, Avastin ^tM, azacitidine, azathioprine, spit of fland, nitrogen Lars, aziridines, basiliximab, BAYX antibody, Bei Laxipu, Baily monoclonal antibody, Belotecan, bendamustine, cypress for wooden monoclonal antibody, bexarotene, bisantrene, bismuth-213 ( ²¹³bi), bismuth-212 ( ²¹²bi), bleomycin, bleomycin, bleomycin, BLyS antibody, Velcade, busulfan, busulfan, Calcineurin inhibitors, calicheamicin, camptothecine, camptothecin, capecitabine, carboplatin (Paraplatin), carboquone, carminomycin, carmofur, carmustine, carmustine (BSNU), CAT antibody, CD11a antibody, CD147/ fundamental immunity globulin antibody, CD154 antibody, CD18 antibody, CD20 antibody, CD23 antibody, CD3 antibody, CD4 antibody, CD40 antibody, CD62L/L-selects plain antibody, CD80 antibody, CDK inhibitor, cedelizumab, celecoxib, Pegylation match trastuzumab, Chlorambucil, Chlorambucil class, S-Neoral, cis-dichlorodiamine platinum (II) (DDP) cis-platinum, CldAdo, clenoliximab, Clofarex, colchicine, complement component 5 antibody, copper-67 (67Cu), reflunomide, CTLA-4 antibody, CTLA-4 fused protein, cyclophilin inhibitor, endoxan, epithio phosphamide, cytosine arabinoside, cytosine arabinoside, cytochalasin B, cytotoxicity rnase, Dacarbazine, daclizumab, gengshengmeisu, gengshengmeisu (dactinomycin), daunomycin, daunomycin, daunomycin (claiming daunorubicin in the past), Decitabine, De Fulimosi, Omaine, detorubicin, mitobronitol, diethylcarbamazine, dihydrofolate reductase inhibitor, dihydroxyl anthracin diketone, diphtheria toxin, archaeal dna polymerase inhibitor, Docetaxel, Dorlimomab Aritox, to get profit western pearl monoclonal antibody, Dx (Zorubicin), DXL625, according to storehouse pearl monoclonal antibody, efalizumab, second method former times sieve, EGFR antagonist, department of Erie is not, elsamitrucin, Ai Ximo monoclonal antibody, Hemometine, endothelin-receptor antagonists, epipodophyllotoxin class, epirubicin, ebormycine class, Erbitux ^tM, the sharp pearl monoclonal antibody of strategic point, Emcyt, etanercept, ethidium bromide, etoglucid, Etoposide, etoposide phosphate, everolimus, faralimomab, farnesyl tranfering enzyme inhibitor, FKBP inhibitor, floxuridine, fludarabine, Fluracil, virtue trastuzumab, fotemustine, galiximab, gallium-67 (67Ga), Ge Telu monoclonal antibody, Jia Weimo monoclonal antibody, gemcitabine, glucocorticoids, the wooden monoclonal antibody of dagger-axe profit, Shandong former times monoclonal antibody, Gramicidin D, gusperimus, hydrazine class, hydroxyurea, hypomethylation agent, idarubicin, idarubicin, ifosfamide, IL-1 antagonist, IL-1 receptor antagonist, IL-12, IL-12 antibody, IL-12R antagonist, IL-13 antibody, IL-2, IL-2 inhibitor, IL-2 acceptor/CD25 antibody, IL-6 antibody, imatinib mesylate, immunoglobulin E antibodies, IMP dehydrogenase inhibitor, infliximab, Inolimomab, integrin antibody, interferon antibody, Interferon, rabbit, interleukin-15 antibody, interleukin-6 receptor antibody, interleukin, iodine-125 ( ¹²⁵i), iodine-131 ( ¹³¹i), her monoclonal antibody, irinotecan, ipsapirone, keliximab, La Luotasai, lead-212 ( ²¹²pb), Lai Buli pearl monoclonal antibody, leflunomide, Revlimid, lerdelimumab, formyl tetrahydrofolic acid, LFA-I antibody, lignocaine, lipoxidase inhibitor, lomustine (CCNU), lonidamine, lucanthone, Shandong former times monoclonal antibody, lutetium-177 ( ¹⁷⁷lu), Macrolide, Mannosulfan, Maslimomab, masoprocol, mechlorethamine, melphalan, mepolizumab, purinethol, beautiful for wooden monoclonal antibody, methotrexate, microtubules inhibitor, microtubule stability toughener, mithramycin, mitobronitol, mitoguazone, mitomycin, ametycin, mitotane, mitoxantrone, morolimumab, mTOR inhibitors, Muromondb-CD3, nitrogen mustards, mycophenolic acid, mitotane (O, P'-(DDD)), natalizumab, S 254, nerelimomab, nimustine, nitrogen mustards, nitrosourea, positive dihydroguaiaretic acid, Ao Limosen, auspicious pearl monoclonal antibody difficult to understand, auspicious pearl monoclonal antibody difficult to understand, Odulimomab, method wood monoclonal antibody difficult to understand, Aura handkerchief Buddhist nun, omalizumab, Ao Tasai, Austria's former times pearl monoclonal antibody, oxaliplatin, oxaliplatin, taxol (PTX), pearl monoclonal antibody examined by handkerchief, PDGF antagonist, pegaspargase, pemetrexed, spray department statin, handkerchief trastuzumab, training gram pearl monoclonal antibody, phosphodiesterase inhibitor, phosphorus-32 ( ³²p), pimecrolimus abetimus, pirarubicin, China fir fine jade, platinum class, Plicamycin, Poly ADP-ribose polymerase inhibitor, porfimer sodium, derivatives of porphyrin, PM, PROCAINE HCL, PHARMA GRADE, Procarbazine, Procarbazine, Proprasylyte, proteasome inhibitor, Pseudomonas exotoxin, pseudomonal toxin, purine synthetic inhibitor, tetracycline, pyrimidine synthesis inhibitors, radionuclide, radiotherapy dose, Raltitrexed, ranomustine, Rayleigh pearl monoclonal antibody, retinoid X receptor agonist, class xanthoplane, rhenium-186 ( ¹⁸⁶re), rhenium-188 ( ¹⁸⁸re), ribonucleotide reductase inhibitors, Ricin, Li Naxipu, rovelizumab, Rubitecan, ruplizumab, samarium-153 ( ¹⁵³sm), Satraplatin, scandium-47 ( ⁴⁷sc), SARM, selective estrogen receptor modulators, fill in sharp Seeley, semustine, sexual hormoue antagonist, cedelizumab, sirolimus, steroidal aromatase inhibitor, steroid, streptozocin, U-9889, tacrolimus, talaporfin, his sharp pearl monoclonal antibody, taxanes, PTX class, Ftorafur, telimomab aritox, temoporfin, Temozolomide, CCI-779, CCI-779, for how former times monoclonal antibody, teniposide, for sharp pearl monoclonal antibody, Teriflunomide, for Si Tasai, testolactone, tetracaine, thalidomide, thiophene is for sending Chlorambucil, thio-purine Tioguanine, thiophene is for group, thymidylate synthetase inhibitor, tiazofurine, for pyrrole method Buddhist nun, T-lymphocyte antibody, TNF antagonist, TNF antibody, TNF fused protein, TNF receptor fusion protein matter, TNF-alpha inhibitor, trastuzumab, topoisomerase enzyme inhibitor, topotecan, hold in the palm sharp pearl monoclonal antibody, ET-743, Sibutramine Hydrochloride wood monoclonal antibody, Treosulfan, vitamin A acid, triazene, triaziquone, Tretamine, four nitric acid three platinum, trofosfamide, specific for tumour antigen monoclonal antibody, tyrosine kinase inhibitor, Uramustine, excellent spy gram monoclonal antibody, valrubicin, valrubicin, cut down profit former times monoclonal antibody, VEGF antagonist, vepalimomab, Visudyne, vinealeucoblastine(VLB), vinca alkaloids, vincristine(VCR), vindesine, Vinflunine, vinorelbine, tie up western pearl monoclonal antibody, Vorinostat, yttrium-88 ( ⁸⁸y), Yttrium-90 ( ⁹⁰y), prick wooden monoclonal antibody, zileuton, draw wooden monoclonal antibody, zolimomab aritox, zorubicin, Zuo Tamosi or its any combination together, and/or

(18) it comprises another promoting agent, and another promoting agent described is comprise the agonist of the following factor, antagonist or conditioning agent: TNF-α, IL-2, IL-4, IL-6, IL-10, IL-12, IL-13, IL-18, iFN-γ, BAFF, CXCL13, IP-10, VEGF, EPO, EGF, HRG, pHGF (HGF), iron adjust element or its any combination.

7. the anti-human HGF antibody according to any one of claim 1 to 6 or fragment or composition, described antibody or fragment:

(1) body internal specific be bonded to express HGF people's cell and/or to circulation solubility HGF molecule;

(2) specific binding is to the HGF on people's cell of expressing in the patient of the relevant disease of the cell suffered to express HGF or the HGF by described people's cell expressing, described diseases such as cancer, comprises ovarian cancer, mammary cancer, lung cancer (minicell or non-small cell), colon and colorectal carcinoma, prostate cancer, carcinoma of the pancreas, kidney, cancer of the stomach, liver cancer, bladder cancer, thyroid carcinoma, carcinoma of endometrium, tumor of head and neck, melanoma, sarcoma, leukemia; Lymphoma; With the brain tumor (such as glioblastoma multiforme) of children or adult; Macular degeneration; Alzheimer; And malaria infection;

(3) contained described V in described antibody _hor V _lpolypeptide originates from one or more rabbit B cell colony;

(4) described antibody or fragment do not have the binding specificity for HGFR; And/or

(5) described antibody or fragment suppress generation and/or the polymeric generation of HGF/HGFR of HGF/HGFR mixture.

8. one or more separated nucleic acid, its encode anti-human HGF antibody or antibody fragment or its variable heavy chain or light chain and express in the host cell be applicable to time cause it to express, described antibody or antibody fragment are selected from:

(1) combined by epitope competition identical or overlapping with the anti-human HGF antibody be selected from the group that is made up of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28 on the described antibody of one or more encoded by nucleic acid described or antibody fragment and people HGF and/or specific binding to described epi-position;

(2) by the antibody of the described expression of one or more encoded by nucleic acid described or antibody fragment specific binding to and people HGF on and be selected from by Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, the epi-position that anti-human HGF antibody in the group that Ab27 with Ab28 forms is identical or overlapping, wherein said epi-position optionally can be differentiated through combining mensuration, described combination measures the combination of one or more peptides in the library detecting anti-human HGF antibody and 10-15-mer peptide, described 10-15-mer peptide is correspond to overlapping fragments that is all or all described people HGF polypeptide length substantially, wherein said combination measures optionally further for protein immunoblotting measures, its specific binding by using chemiluminescent labeling to detect described antibody or antibody fragment and 10-15mer peptide described in one or more in described library, launches detectable chemiluminescence signal during the peptide of described chemiluminescent labeling in described antibody or antibody fragment specific binding extremely described library,

(3) contain by the antibody of the described expression of one or more encoded by nucleic acid described or antibody fragment at least 2 complementary determining regions (CDR) be selected from by the anti-HGF antibody in following formed group: Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28;

(4) contain by the anti-human anti-HGF antibody of the described expression of one or more encoded by nucleic acid described or antibody fragment at least 3 complementary determining regions (CDR) be selected from by the anti-HGF antibody in following formed group: Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28;

(5) contain by the anti-human HGF antibody of the described expression of one or more encoded by nucleic acid described or antibody fragment at least 4 complementary determining regions (CDR) be selected from by the anti-HGF antibody in following formed group: Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28;

(6) contain by the anti-human HGF antibody of the described expression of one or more encoded by nucleic acid described or antibody fragment at least 5 complementary determining regions (CDR) be selected from by the anti-HGF antibody in following formed group: Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28;

(7) contain by the antibody of the described expression of one or more encoded by nucleic acid described or antibody fragment whole 6 complementary determining regions (CDR) be selected from by the anti-HGF antibody in following formed group: Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab23, Ab24, Ab25, Ab26, Ab27 and Ab28;

(8) contain (a) variable heavy chain by the anti-human HGF antibody of the described expression of one or more encoded by nucleic acid described or antibody fragment, it comprises the CDR1 sequence be selected from by following formed group:

(a) SEQIDNO:4, SEQIDNO:44, SEQIDNO:84, SEQIDNO:124, SEQIDNO:164, SEQIDNO:204, SEQIDNO:244, SEQIDNO:284, SEQIDNO:324, SEQIDNO:364, SEQIDNO:404, SEQIDNO:444, SEQIDNO:484, SEQIDNO:524, SEQIDNO:564, SEQIDNO:604, SEQIDNO:644, SEQIDNO:684, SEQIDNO:724, SEQIDNO:764, SEQIDNO:804, SEQIDNO:844, SEQIDNO:884 and SEQIDNO:924, SEQIDNO:964, SEQIDNO:1004 and SEQIDNO:1044, be selected from by SEQIDNO:6, SEQIDNO:46, SEQIDNO:86, SEQIDNO:126, SEQIDNO:166, SEQIDNO:206, SEQIDNO:246, SEQIDNO:286, SEQIDNO:326, SEQIDNO:366, SEQIDNO:406, SEQIDNO:446, SEQIDNO:486, SEQIDNO:526, SEQIDNO:566, SEQIDNO:606, SEQIDNO:646, SEQIDNO:686, SEQIDNO:726, SEQIDNO:766, SEQIDNO:806, SEQIDNO:846, SEQIDNO:886 and SEQIDNO:926, SEQIDNO:966, CDR2 sequence in the group that SEQIDNO:1006 and SEQIDNO:1046 forms, with be selected from by SEQIDNO:8, SEQIDNO:48, SEQIDNO:88, SEQIDNO:128, SEQIDNO:168, SEQIDNO:208, SEQIDNO:248, SEQIDNO:288, SEQIDNO:328, SEQIDNO:368, SEQIDNO:408, SEQIDNO:448, SEQIDNO:488, SEQIDNO:528, SEQIDNO:568, SEQIDNO:608, SEQIDNO:648, SEQIDNO:688, SEQIDNO:728, SEQIDNO:768, SEQIDNO:808, SEQIDNO:848, SEQIDNO:888 and SEQIDNO:928, SEQIDNO:968, CDR3 sequence in the group that SEQIDNO:1008 and SEQIDNO:1048 forms, and/or

(b) variable light, it comprises and is selected from by SEQIDNO:24, SEQIDNO:64, SEQIDNO:104, SEQIDNO:144, SEQIDNO:184, SEQIDNO:224, SEQIDNO:264, SEQIDNO:304, SEQIDNO:344, SEQIDNO:384, SEQIDNO:424, SEQIDNO:464, SEQIDNO:504, SEQIDNO:544, SEQIDNO:584, SEQIDNO:624, SEQIDNO:664, SEQIDNO:704, SEQIDNO:744, SEQIDNO:784, SEQIDNO:824, SEQIDNO:864, SEQIDNO:904 and SEQIDNO:944, SEQIDNO:984, CDR1 sequence in the group that SEQIDNO:1024 and SEQIDNO:1064 forms, be selected from by SEQIDNO:26, SEQIDNO:66, SEQIDNO:106, SEQIDNO:146, SEQIDNO:186, SEQIDNO:226, SEQIDNO:266, SEQIDNO:306, SEQIDNO:346, SEQIDNO:386, SEQIDNO:426, SEQIDNO:466, SEQIDNO:506, SEQIDNO:546, SEQIDNO:586, SEQIDNO:626, SEQIDNO:666, SEQIDNO:706, SEQIDNO:746, SEQIDNO:786, SEQIDNO:826, SEQIDNO:866, SEQIDNO:906 and SEQIDNO:946, SEQIDNO:986, CDR2 sequence in the group that SEQIDNO:1026 and SEQIDNO:1066 forms, with be selected from by SEQIDNO:28, SEQIDNO:68, SEQIDNO:108, SEQIDNO:148, SEQIDNO:188, SEQIDNO:228, SEQIDNO:268, SEQIDNO:308, SEQIDNO:348, SEQIDNO:388, SEQIDNO:428, SEQIDNO:468, SEQIDNO:508, SEQIDNO:548, SEQIDNO:588, SEQIDNO:628, SEQIDNO:668, SEQIDNO:708, SEQIDNO:748, SEQIDNO:788, SEQIDNO:828, SEQIDNO:868, SEQIDNO:908, SEQIDNO:948, SEQIDNO:988, CDR3 sequence in the group that SEQIDNO:1028 and SEQIDNO:1068 forms, its further restricted condition for one or two residue of arbitrary previously differentiated CDR polypeptide can through another aminoacid replacement,

(9) the CDR3 sequence of the CDR1 sequence of SEQIDNO:4, the CDR2 sequence of SEQIDNO:6 and SEQIDNO:8 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:24, the CDR2 sequence of SEQIDNO:26 and SEQIDNO:28;

(10) the CDR3 sequence of the CDR1 sequence of SEQIDNO:44, the CDR2 sequence of SEQIDNO:46 and SEQIDNO:48 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:64, the CDR2 sequence of SEQIDNO:66 and SEQIDNO:68;

(11) the CDR3 sequence of the CDR1 sequence of SEQIDNO:84, the CDR2 sequence of SEQIDNO:86 and SEQIDNO:88 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:104, the CDR2 sequence of SEQIDNO:106 and SEQIDNO:108;

(12) the CDR3 sequence of the CDR1 sequence of SEQIDNO:124, the CDR2 sequence of SEQIDNO:126 and SEQIDNO:128 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:144, the CDR2 sequence of SEQIDNO:146 and SEQIDNO:148;

(13) the CDR3 sequence of the CDR1 sequence of SEQIDNO:164, the CDR2 sequence of SEQIDNO:166 and SEQIDNO:168 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:184, the CDR2 sequence of SEQIDNO:186 and SEQIDNO:188;

(14) the anti-human HGF antibody of described expression or antibody fragment contain variable heavy chain, and it comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:204, the CDR2 sequence of SEQIDNO:206 and SEQIDNO:208; And described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:224, the CDR2 sequence of SEQIDNO:226 and SEQIDNO:228;

(15) the CDR3 sequence of the CDR1 sequence of SEQIDNO:244, the CDR2 sequence of SEQIDNO:246 and SEQIDNO:248 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:264, the CDR2 sequence of SEQIDNO:266 and SEQIDNO:268;

(16) the CDR3 sequence of the CDR1 sequence of SEQIDNO:284, the CDR2 sequence of SEQIDNO:286 and SEQIDNO:288 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:304, the CDR2 sequence of SEQIDNO:306 and SEQIDNO:308;

(17) the CDR3 sequence of the CDR1 sequence of SEQIDNO:324, the CDR2 sequence of SEQIDNO:326 and SEQIDNO:328 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:344, the CDR2 sequence of SEQIDNO:346 and SEQIDNO:348;

(18) the CDR3 sequence of the CDR1 sequence of SEQIDNO:364, the CDR2 sequence of SEQIDNO:366 and SEQIDNO:368 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:384, the CDR2 sequence of SEQIDNO:386 and SEQIDNO:388;

(19) the CDR3 sequence of the CDR1 sequence of SEQIDNO:404, the CDR2 sequence of SEQIDNO:406 and SEQIDNO:408 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:424, the CDR2 sequence of SEQIDNO:426 and SEQIDNO:428;

(20) the CDR3 sequence of the CDR1 sequence of SEQIDNO:444, the CDR2 sequence of SEQIDNO:446 and SEQIDNO:448 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:464, the CDR2 sequence of SEQIDNO:466 and SEQIDNO:468;

(21) the CDR3 sequence of the CDR1 sequence of SEQIDNO:484, the CDR2 sequence of SEQIDNO:486 and SEQIDNO:488 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:504, the CDR2 sequence of SEQIDNO:506 and SEQIDNO:508;

(22) the CDR3 sequence of the CDR1 sequence of SEQIDNO:524, the CDR2 sequence of SEQIDNO:526 and SEQIDNO:528 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:544, the CDR2 sequence of SEQIDNO:546 and SEQIDNO:548;

(23) the CDR3 sequence of the CDR1 sequence of SEQIDNO:564, the CDR2 sequence of SEQIDNO:566 and SEQIDNO:568 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:584, the CDR2 sequence of SEQIDNO:586 and SEQIDNO:588;

(24) the CDR3 sequence of the CDR1 sequence of SEQIDNO:604, the CDR2 sequence of SEQIDNO:606 and SEQIDNO:608 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:624, the CDR2 sequence of SEQIDNO:626 and SEQIDNO:628;

(25) the CDR3 sequence of the CDR1 sequence of SEQIDNO:644, the CDR2 sequence of SEQIDNO:646 and SEQIDNO:648 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:664, the CDR2 sequence of SEQIDNO:666 and SEQIDNO:668;

(26) the CDR3 sequence of the CDR1 sequence of SEQIDNO:684, the CDR2 sequence of SEQIDNO:686 and SEQIDNO:688 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:704, the CDR2 sequence of SEQIDNO:706 and SEQIDNO:708;

(27) the CDR3 sequence of the CDR1 sequence of SEQIDNO:724, the CDR2 sequence of SEQIDNO:726 and SEQIDNO:728 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:744, the CDR2 sequence of SEQIDNO:746 and SEQIDNO:748;

(28) the CDR3 sequence of the CDR1 sequence of SEQIDNO:764, the CDR2 sequence of SEQIDNO:766 and SEQIDNO:768 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:784, the CDR2 sequence of SEQIDNO:786 and SEQIDNO:788;

(29) the CDR3 sequence of the CDR1 sequence of SEQIDNO:804, the CDR2 sequence of SEQIDNO:806 and SEQIDNO:808 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:824, the CDR2 sequence of SEQIDNO:826 and SEQIDNO:828;

(30) the CDR3 sequence of the CDR1 sequence of SEQIDNO:844, the CDR2 sequence of SEQIDNO:846 and SEQIDNO:848 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or described variable light comprises the CDR3 sequence of the CDR1 sequence of SEQIDNO:864, the CDR2 sequence of SEQIDNO:866 and SEQIDNO:868;

(31) the CDR3 sequence of the CDR1 sequence of SEQIDNO:884, the CDR2 sequence of SEQIDNO:886 and SEQIDNO:888 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or the CDR3 sequence of the CDR1 sequence of SEQIDNO:904, the CDR2 sequence of SEQIDNO:906 and SEQIDNO:908 is comprised by the described variable light of one or more encoded by nucleic acid described;

(32) the CDR3 sequence of the CDR1 sequence of SEQIDNO:924, the CDR2 sequence of SEQIDNO:926 and SEQIDNO:928 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or the CDR3 sequence of the CDR1 sequence of SEQIDNO:944, the CDR2 sequence of SEQIDNO:946 and SEQIDNO:948 is comprised by the described variable light of one or more encoded by nucleic acid described;

(33) the CDR3 sequence of the CDR1 sequence of SEQIDNO:964, the CDR2 sequence of SEQIDNO:966 and SEQIDNO:968 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or the CDR3 sequence of the CDR1 sequence of SEQIDNO:984, the CDR2 sequence of SEQIDNO:986 and SEQIDNO:988 is comprised by the described variable light of one or more encoded by nucleic acid described;

(34) the CDR3 sequence of the CDR1 sequence of SEQIDNO:1004, the CDR2 sequence of SEQIDNO:1006 and SEQIDNO:1008 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or the CDR3 sequence of the CDR1 sequence of SEQIDNO:1024, the CDR2 sequence of SEQIDNO:1026 and SEQIDNO:1028 is comprised by the described variable light of one or more encoded by nucleic acid described;

(35) the CDR3 sequence of the CDR1 sequence of SEQIDNO:1044, the CDR2 sequence of SEQIDNO:1046 and SEQIDNO:1048 is comprised by the described variable heavy chain of one or more encoded by nucleic acid described; And/or the CDR3 sequence of the CDR1 sequence of SEQIDNO:1064, the CDR2 sequence of SEQIDNO:1066 and SEQIDNO:1068 is comprised by the described variable light of one or more encoded by nucleic acid described;

(36) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:2 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence; And/or comprise SEQIDNO:22 by the described variable light of one or more encoded by nucleic acid described;

(37) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:2 with SEQIDNO:22 respectively separately with light chain by the described variable heavy chain of described encoded by nucleic acid;

(38) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:42 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:62 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(39) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:42 with SEQIDNO:62 respectively separately with light chain by the described variable heavy chain of described encoded by nucleic acid;

(40) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:82 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:102 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(41) described variable heavy chain and light chain are identical with light chain at least 90% with the variable heavy chain in SEQIDNO:82 with SEQIDNO:102 respectively separately;

(42) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:122 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:142 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(43) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:122 with SEQIDNO:142 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(44) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:162 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:182 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(45) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:162 with SEQIDNO:182 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(46) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:202 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence; And by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:222 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(47) described variable heavy chain and light chain are identical with light chain at least 90% with the variable heavy chain in SEQIDNO:202 with SEQIDNO:222 respectively separately;

(48) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:242 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:262 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(49) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:242 with SEQIDNO:262 respectively separately with light chain by the described variable heavy chain of described encoded by nucleic acid;

(50) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:282 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:302 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(51) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:282 with SEQIDNO:302 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(52) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:322 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:342 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(53) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:322 with SEQIDNO:342 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(54) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:362 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:382 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(55) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:362 with SEQIDNO:382 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(56) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:402 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:422 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(57) described variable heavy chain and light chain are identical with light chain at least 90% with the variable heavy chain in SEQIDNO:402 with SEQIDNO:422 respectively separately;

(58) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:442 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:462 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(59) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:442 with SEQIDNO:462 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(60) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:482 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:502 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(61) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:482 with SEQIDNO:502 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(62) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:522 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:542 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(63) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:522 with SEQIDNO:542 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(64) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:562 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:582 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(65) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:562 with SEQIDNO:582 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(66) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:602 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:622 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(67) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:602 with SEQIDNO:622 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(68) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:642 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence; And by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:662 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(69) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:642 with SEQIDNO:662 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(70) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:682 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:702 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(71) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:682 with SEQIDNO:702 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(72) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:722 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:742 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(73) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:722 with SEQIDNO:742 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(74) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:762 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence; And by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:782 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(75) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:762 with SEQIDNO:782 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(76) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:802 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:822 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(77) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:802 with SEQIDNO:822 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(78) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:842 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:862 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(79) described variable heavy chain and light chain are identical with light chain at least 90% with the variable heavy chain in SEQIDNO:842 with SEQIDNO:862 respectively separately;

(80) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:882 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:902 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(81) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:882 with SEQIDNO:902 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(82) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:922 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:942 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(83) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:922 with SEQIDNO:942 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(84) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:962 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:982 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(85) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:962 with SEQIDNO:982 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(86) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:1002 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:1022 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(87) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:1002 with SEQIDNO:1022 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(88) by the described variable heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:1042 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described variable light of one or more encoded by nucleic acid described comprise with SEQIDNO:1062 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(89) identical with light chain at least 90% with the variable heavy chain in SEQIDNO:1042 with SEQIDNO:1062 respectively separately with light chain by the described variable heavy chain of one or more encoded by nucleic acid described;

(90) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:1 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:21 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(91) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:41 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:61 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(92) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:81 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:101 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(93) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:121 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:141 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(94) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:161 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:181 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(95) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:201 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:221 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(96) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:241 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:261 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(97) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:281 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:301 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(98) comprise SEQIDNO:321 by the described heavy chain of one or more encoded by nucleic acid described, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:341 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(99) by the heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:361 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and comprise SEQIDNO:381 by the described light chain of one or more encoded by nucleic acid described;

(100) comprise SEQIDNO:401 by the described heavy chain of one or more encoded by nucleic acid described, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:421 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(101) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:441 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:461 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(102) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:481 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:501 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(103) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:521 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and comprise SEQIDNO:541 by the described light chain of one or more encoded by nucleic acid described;

(104) comprise SEQIDNO:561 by the described heavy chain of one or more encoded by nucleic acid described, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:581 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(105) by one or more encoded by nucleic acid described described heavy chain and SEQIDNO:601 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and comprise SEQIDNO:621 by the described light chain of one or more encoded by nucleic acid described;

(106) comprise SEQIDNO:641 by the described heavy chain of one or more encoded by nucleic acid described, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:661 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(107) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:681 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:701 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(108) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:721 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and comprise SEQIDNO:741 by the described light chain of one or more encoded by nucleic acid described;

(109) comprise SEQIDNO:761 by the described heavy chain of one or more encoded by nucleic acid described, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:781 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(110) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:801 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:821 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(111) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:841 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:861 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(112) by the described heavy chain of one or more encoded by nucleic acid described comprise with SEQIDNO:881 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence, and comprise SEQIDNO:901 by the described light chain of one or more encoded by nucleic acid described;

(113) comprise SEQIDNO:921 by the described heavy chain of one or more encoded by nucleic acid described, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:941 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(114) comprise SEQIDNO:961 by the described heavy chain of one or more encoded by nucleic acid described, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:981 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence;

(115) comprise SEQIDNO:1001 by the described heavy chain of one or more encoded by nucleic acid described, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:1021 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence; And/or

(116) comprise SEQIDNO:1041 by the described heavy chain of one or more encoded by nucleic acid described, and by the described light chain of one or more encoded by nucleic acid described comprise with SEQIDNO:1061 at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical sequence; Or any combination of above-mentioned each.

9. one or more nucleic acid as claimed in claim 8, are wherein selected from by the described antibody of one or more encoded by nucleic acid described or antibody fragment:

(1) chimeric, humanization and people's antibody or antibody fragment;

(2) scFv, camel antibodies, nano antibody, IgNAR, Fab fragment, Fab' fragment, MetMab, monovalent antibody fragments and F (ab') ₂fragment;

(3) N-glycosylation and/or O-glycosylation is lacked substantially or completely by the antibody of the described expression of one or more encoded by nucleic acid described or antibody fragment;

(4) people's constant domain is comprised by the antibody of the described expression of one or more encoded by nucleic acid described or antibody fragment;

(5) be IgG1, IgG2, IgG3 or IgG4 antibody by the antibody of the described expression of one or more encoded by nucleic acid described;

(6) one or more nucleic acid described comprise coding and SEQIDNO:12, SEQIDNO:32, SEQIDNO:52, SEQIDNO:72, SEQIDNO:92, SEQIDNO:112, SEQIDNO:132, SEQIDNO:152, SEQIDNO:172, SEQIDNO:192, SEQIDNO:212, SEQIDNO:232, SEQIDNO:252, SEQIDNO:272, SEQIDNO:292, SEQIDNO:312, SEQIDNO:332, SEQIDNO:352, SEQIDNO:372, SEQIDNO:392, SEQIDNO:412, SEQIDNO:432, SEQIDNO:452, SEQIDNO:472, SEQIDNO:492, SEQIDNO:512, SEQIDNO:532, SEQIDNO:552, SEQIDNO:572, SEQIDNO:592, SEQIDNO:612, SEQIDNO:632, SEQIDNO:652, SEQIDNO:672, SEQIDNO:692, SEQIDNO:712, SEQIDNO:732, SEQIDNO:752, SEQIDNO:772, SEQIDNO:792, SEQIDNO:812, SEQIDNO:832, SEQIDNO:852, SEQIDNO:872, SEQIDNO:892, SEQIDNO:912, SEQIDNO:932, SEQIDNO:952, SEQIDNO:972, SEQIDNO:992, SEQIDNO:1012, SEQIDNO:1032, SEQIDNO:1052 and SEQIDNO:1072 at least 80%, 90%, 95%, 96%, 97%, 98%, the sequence in 99% or 100% identical VH and VL district, or any one codon degeneracy above-mentioned, and/or

(7) one or more nucleotide sequences described comprise Pichia or people's preferred codons.

10. one or more carriers, it comprises one or more nucleotide sequences as described in any of claims 8 or 9.

11. 1 kinds of host cells, it comprises one or more nucleotide sequences as described in any of claims 8 or 9, or one or more carriers as claimed in claim 10.

12. host cells as claimed in claim 11, wherein said host cell is Mammals, bacterium, fungi, yeast, bird or insect cell.

13. host cells as claimed in claim 12, it comprises:

(1) filamentous fungus or yeast;

(2) filamentous fungus or yeast, it is selected from pichia pastoris phaff, Finland's pichia spp, happiness trehalose pichia spp, koala nurse pichia spp, Pichia membranaefaciens, (small Europe adds iron bacteria to small pichia spp, Lin Shi pichia spp), Root and stem of Cholla pichia spp, heat-resisting pichia spp, Spiked Loosestrife pichia spp, pine oak pichia spp, Pi Jiepushi pichia spp, tool handle pichia spp, pichia methanolica, Pichia certain, yeast saccharomyces cerevisiae, yeast belong certain, multiple-shaped nuohan inferior yeast, genus kluyveromyces certain, Kluyveromyces lactis, Candida albicans, little nido aspergillus, black aspergillus, rice aspergillus, Trichodermareesei, Lu Kenuo train of thought gold pityrosporion ovale, Fusarium certain, Fusarium graminearum, fusarium, small liwan moss Neuraspora crassa, Pichia certain, any yeast belong certain, multiple-shaped nuohan inferior yeast, any genus kluyveromyces certain, Candida albicans, any aspergillus certain, Trichodermareesei, Lu Kenuo train of thought gold pityrosporion ovale, any Fusarium certain and Neuraspora crassa,

(3) mammalian cell; And/or

(4) CHO, COS, BHK, myelomatosis, monkey kidney CV1 system (COS-7, ATCCCRL1651) that transformed by SV40; Human embryo kidney (HEK) system (through subclone for grow in suspension culture 293 or 293 cells), mouse C1-esteraseremmer-N profit cell; Human pneumonocyte; Human liver cell or mouse mammary tumor cells.

Prepare the method for antibody according to any one of Claims 1-4 or antibody fragment by express nucleic acid for 14. 1 kinds, described nucleic acid encoding to express described antibody or antibody fragment in recombinant host cell.

15. methods as claimed in claim 14, wherein said host cell is selected from:

(1) bacterium, yeast, fungi, insect cell, vegetable cell, avian cells or mammalian cell;

(2) yeast or filamentous fungus, it is optionally polyploid;

(3) yeast or fungi, it is selected from pichia pastoris phaff, Finland's pichia spp, happiness trehalose pichia spp, koala nurse pichia spp, Pichia membranaefaciens, (small Europe adds iron bacteria to small pichia spp, Lin Shi pichia spp), Root and stem of Cholla pichia spp, heat-resisting pichia spp, Spiked Loosestrife pichia spp, pine oak pichia spp, Pi Jiepushi pichia spp, tool handle pichia spp, pichia methanolica, Pichia certain, yeast saccharomyces cerevisiae, yeast belong certain, multiple-shaped nuohan inferior yeast, genus kluyveromyces certain, Kluyveromyces lactis, Candida albicans, little nido aspergillus, black aspergillus, rice aspergillus, Trichodermareesei, Lu Kenuo train of thought gold pityrosporion ovale, Fusarium certain, Fusarium graminearum, fusarium, small liwan moss and Neuraspora crassa, Pichia certain, any yeast belong certain, multiple-shaped nuohan inferior yeast, any genus kluyveromyces certain, Candida albicans, any aspergillus certain, Trichodermareesei, Lu Kenuo train of thought gold pityrosporion ovale, any Fusarium certain and Neuraspora crassa,

(4) mammalian cell;

(5) CHO, COS, BHK, myelomatosis, monkey kidney CV1 system (COS-7, ATCCCRL1651) that transformed by SV40; Human embryo kidney (HEK) system (through subclone for grow in suspension culture 293 or 293 cells), mouse C1-esteraseremmer-N profit cell; Human pneumonocyte; Human liver cell or mouse mammary tumor cells; Or

(6) Chinese hamster ovary celI.

16. methods as claimed in claim 14, wherein said recombinant host cell is stably express at least 10-25mg/ rise described antibody or antibody fragment and secreted to the monoploid in substratum or polyploid yeast culture.

17. methods as claimed in claim 16, wherein said polyploid yeast is by comprising following method preparation:

I at least one expression vector is introduced in haploid yeast cell by (), described expression vector contains one or more heterologous polynucleotide of encoding said antibody, and it may be operably coupled to promotor and signal sequence;

(ii) polyploid yeast is produced by mating or spheraplast fusion from the described first and/or second haploid yeast cell;

(iii) the polyploid yeast cell of stably expressing described antibody is selected; With

(iv) the described polyploid yeast cell that self stabilization ground is expressed in described antibody to described substratum produces stable polyploid yeast culture.

18. methods as claimed in claim 17, wherein said yeast belong is Pichia.

19. 1 kinds of treatments or diagnostic method, wherein said method comprises uses anti-HGF antibody or antibody fragment, and described method is selected from following:

(1) to the patient therapeuticallv of the relevant disease of the process LAN suffered to express the cell of HGF or HGF or symptom or prevention or diagnose the anti-human HGF antibody of at least one according to any one of claim 1 to 7 of significant quantity or fragment or contain its composition;

(2) adjust the vasculogenesis that HGF in (suppress, block or promote) experimenter in need is relevant, it comprises the anti-human HGF antibody of at least one according to any one of claim 1 to 7 of administering therapeutic or prevention significant quantity or fragment or the composition containing it;

(3) adjust the cell proliferation that HGF in (suppress, block or promote) experimenter in need is relevant, it comprises the anti-human HGF antibody of at least one according to any one of claim 1 to 7 of administering therapeutic or prevention significant quantity or fragment or the composition containing it;

(4) adjust the cell invasion that HGF in (suppress, block or promote) experimenter in need is relevant, it comprises the anti-human HGF antibody of at least one according to any one of claim 1 to 7 of administering therapeutic or prevention significant quantity or fragment or the composition containing it;

(5) suppress or block the relevant transfer of HGF in experimenter in need, it comprises the anti-human HGF antibody of at least one according to any one of claim 1 to 7 of administering therapeutic or prevention significant quantity or fragment or the composition containing it;

(6) adjust HGF/HGF-R (c-met) in (suppress, block or promote) experimenter in need mutually mutually with and/or c-met activation, it comprises the anti-human HGF antibody of at least one according to any one of claim 1 to 7 of administering therapeutic or prevention significant quantity or fragment or the composition containing it;

(7) suppress or block the mitogenesis of the HGF in experimenter in need, prokinetic and/or short form forms character, it comprises administering therapeutic or the prevention anti-human HGF antibody of at least one according to any one of claim 1 to 7 of significant quantity or fragment or the composition containing it;

(8) promote the fibrosis that HGF in experimenter in need is relevant, it comprises the anti-human HGF antibody of at least one according to any one of claim 1 to 7 of administering therapeutic or prevention significant quantity or fragment or the composition containing it; And/or

(9) suppress the cell dispersal that HGF in experimenter in need is relevant, it comprises the anti-human HGF antibody of at least one according to any one of claim 1 to 7 of administering therapeutic or prevention significant quantity or fragment or the composition containing it; Or its any combination.

20. methods as claimed in claim 19, wherein

(1) described antibody or fragment suppress or prevent HGF/c-met from combining;

(2) described antibody or fragment do not suppress or prevent HGF/c-met from combining;

(3) described antibody or fragment suppress c-met activation;

(4) described antibody or fragment are bonded to HGF/HGF-R mixture or polymer.

21. methods as described in claim 19 or 20, wherein said method comprises uses another therapeutical agent.

22. methods as claimed in claim 21, another therapeutical agent wherein said is selected from: chemotherapeutic, Statins, cytokine, immunosuppressor, gene therapeutic agents, anti-coagulant, anti-emaciation agent, anti-unable dose, fatigue protective agent, anti-heat-generating agent, nausea agent, antiemetic, IL-6 antagonist, cytotoxic agent, analgesic agent, febrifuge, antiphlogistic, microbiotic, antiviral agent, antibacterial agent agent, anti-angiogenic agent or its any combination.

23. methods as claimed in claim 22, wherein:

(1) described chemotherapeutic is selected from: VEGF antagonist, EGFR antagonist, platinum class, PTX class, irinotecan, 5 FU 5 fluorouracil, gemcitabine, formyl tetrahydrofolic acid, steroid, endoxan, melphalan, vinca alkaloids, vinealeucoblastine(VLB), vincristine(VCR), vindesine, vinorelbine, nitrogen mustards, tyrosine kinase inhibitor, radiotherapy dose, sexual hormoue antagonist, SARM, selective estrogen receptor modulators, PDGF antagonist, TNF antagonist, IL-I antagonist, interleukin, IL-12, IL-2, IL-12R antagonist, the monoclonal antibody that toxin is puted together, specific for tumour antigen monoclonal antibody, Erbitux ^tM, Avastin ^tM, handkerchief trastuzumab, anti-CD20 antibodies, auspicious pearl monoclonal antibody difficult to understand, method difficult to understand wood monoclonal antibody, DXL625, or its any combination,

(2) described anti-coagulant is selected from ReoPro (ReoPro ^tM), acenocoumarol, antithrombin IE, argatroban, acetylsalicylic acid, Bivalirudin (Angiomax ^tM), clopidogrel, dabigatran, dabigatran etcxilate (Pradaxa ^tM/ Pradax ^tM), desirudin (Revasc ^tM/ Iprivask ^tM), Dipyridamole, eptifibatide (Integrilin ^tM), fondaparin, heparin, r-hirudin, according to DALT, Lepirudin (Refludan ^tM), low molecular weight heparin, Melagatran, phenindione, phenprocoumon, ticlopidine, Tirofiban (Aggrastat ^tM), warfarin, ximelagatran, ximelagatran (Exanta ^tM/ Exarta ^tM) or its any combination;

(3) described statin is selected from: atorvastatin, Cerivastatin, fluvastatin, lovastatin, mevastatin, Pravastatin, Pravastatin, Rosuvastatin, Simvastatin or its any combination;

(4) another therapeutical agent described is the antagonist being selected from the following factor: tumor necrosis factor-alpha, interferon-gamma, interleukin-11 α, interleukin-11 β, interleukin 6, proteolysis inducible factor, leukaemia inhibitory factor or its any combination Pichia

(5) another therapeutical agent described is be selected from following anti-angiogenesis: soluble VEGFR-1, NRP-1, ANG2, TSP-1 and TSP-2, angiostatin and associated molecule, endostatin, vascularization statin, calprotectin, PF4, TIMP, CDAI, Meth-1, Meth-2, IFN-α, IFN-β and IFN-γ, CXCL10, IL-4, IL-12 and IL-18, thrombogen (kringle structural domain-2), Antithrombin III fragment, prolactin antagonist, VEGI, SPARC, osteopontin, mammary gland serine protease inhibitor and canstatin,

(6) another therapeutical agent described is comprise following anti-emaciation agent: hemp, dronabinol (Marinol ^tM), nabilone (Cesamet), cannabidiol, cannabichromene, tetrahydrocannabinol, Sativex, megestrol acetate or its any combination;

(7) another therapeutical agent described is nausea agent or antiemetic, and it comprises: 5-HT3 receptor antagonist, asafoetide, alizapride, anticholinergic, antihistamine, Aprepitant, benzodiazepine class, cannabichromene, cannabidiol, cannaboid, hemp, Carcel smooth, chlorpromazine, marezine, dexamethasone, dexamethasone, umine (Gravol ^tM), diphenhydramine, dolasetron, domperidone, dopamine antagonist, doxylamine, dronabinol (Marinol ^tM), droperidol, more tell peaceful, ginger, granisetron, haloperidol, hydroxyzine, the sweet henbane alkali in east, lorazepam, Meclozine, Reglan, midazolam, muscimol, nabilone (Cesamet), nkl receptor antagonist, ondansetron, Palonosetron, Mentha arvensis L. syn.M.haplocalyxBrig, promethazine hydrochloride, prochlorperazine, Promacot, promethazine, Pentazine, Disoprofol, sativex, tetrahydrocannabinol, trimethobenzamide, tropisetron, nandrolone, stilboestrol, thalidomide, Revlimid, Leptin agonist, muscle mass antagonist, anti-muscle mass antibody, SARM, selective estrogen receptor modulators, Angiotensin All antagonist, beta 2 adrenoreceptor agonists, β 3 adrenoceptor agonists, or its any combination,

(8) another therapeutical agent described comprises tamoxifen, BCL-2 antagonist, oestrogenic hormon, diphosphonate, teriparatide, strontium ranelate, alendronate sodium (Fosamax), risedronate (safe and reliable good), raloxifene, ibandronate (Bang Luoli), Ao Bakela, ABT-263, gossypol, Gefitinib, epidermal growth factor recipient tyrosine kinase inhibitor, erlotinib, epidermal growth factor receptor inhibitor, psoralene, neosoralen, soloxsalen, bergapton, class xanthoplane, etretinate, Etretin, infliximab adalimumab, infliximab, etanercept, Zenapax ^tM, S-Neoral, methotrexate, G-CSF, filgrastim, lenograstim, excellent Bao Jin, Niu Lasita, 2-arylpropionic acid, Aceclofenac, acemetacin, acetylsalicylic acid (acetylsalicylic acid), Warner-Lambert), alminoprofen, amoxycilline Trihydrate bp, aminoantipyrene, aryl alkanoic acid, azapropazone, Benorilate/Benorilate, Compd 90459, Bromfenac, carprofen, celecoxib, choline salicylate magnesium, Perclusone, cox 2 inhibitor, Seractil, dexketoprofen, diclofenac, diflunisal, Droxicam, ethenzamide, R-ETODOLAC, Etoricoxib, Fes amine, that acid fragrant, fenbufen, fenoprofen, Flufenamic Acid, flunoxaprofen, flurbiprofen, Ibuprofen BP/EP, ibuproxam, indomethacin, indoprofen, recheton, Ketoprofen BP 93, ketorolac, lornoxicam, loxoprofen, lumiracoxib, magnesium salicylate, meclofenamic acid, mefenamic acid, meloxicam, Sulpyrine, wintergreen oil, mofebutazone, nabumetone, Naproxen Base, the adjacent amido phenylformic acid of N-aryl, oxametacin, Taisho), former times health class, Tacote, parecoxib, phenazone, phenylbutazone, phenylbutazone, piroxicam, pirprofen, profen, proglumetacin, pyrazolidine derivatives, rofecoxib, sasapyrin, salicylic amide, salicylate, sulfinpyrazone, sulindac, sutoprofen, tenoxicam, tiaprofenic acid, tolfenamic acid, tolmetin and valdecoxib, microbiotic, amikacin, aminoglycoside, amoxycilline Trihydrate bp, Ampicillin Trihydrate, ansamycins, Arsphenamine, Azythromycin, azlocillin, aztreonam, bacitracin, carbacephem, carbapenems, Pyocianil, cefaclor, S 578, Cephalexin Monohydrate Micro/Compacted, cefoxitin, cefoxitin, Cefamandole, Cephazolin, Cefdinir, cefditoren, cefepime, Cefixime Micronized, cefoperazone, cefotaxime, cefoxitin, Cefpodoxime, cefprozil, ceftazime, ceftibuten, ceftizoxime, Ceftobiprole, ceftriaxone, cephalofruxin, cephalosporin class, paraxin, cilastatin, Ciprofloxacin, clarithromycin, clindamycin, cloxacillin, Totazina, trimethoprim-sulfamethoxazole, dalfopristin, Demethylchlortetracycline, dicloxacillin, dirithromycin, doripenem, Vibravenos, enoxacin, ertapenem, erythromycin, Tibutol, Flucloxacillin, phosphonomycin, Nifurazolidone, fusidinic acid, Gatifloxacin, geldanamycin, gentamicin, glycopeptide class, herbimycin, imipenum, vazadrine, kantlex, levofloxacin, lincomycin, Linezolid, lomefloxacin, Loracarbef, Macrolide, mafenide, meropenem, X-1497, metronidazole, mezlocillin, MINOCYCLINE HCL, monobactam class, Moxifloxacin, mupirocin, nafcillin, Liu Suanyan NEOMYCIN SULPHATE, netilmicin, furadantin, norfloxicin, Ofloxacine USP 23, Oxazacillin, terramycin, paromycin, penicillin, penicillins, piperacillin, dull and stereotyped mycin, PXB, polypeptide, Prontosil, pyrazinoic acid amide, quinolones, Quinupristin, Rifampin, Rifampin, Roxithromycin, spectinomycin, Streptomycin sulphate, Sulphacetamide, ayerlucil, sulfanilamide (SN), sulfasalazine, Sulfafurazole, sulfamido, teicoplanin, Ketek, tsiklomitsin, tetracyclines, ticarcillin, fasigyne, tobramycin, Trimethoprim BP, Trimethoprim BP-sulfamethoxazole, troleomycin, trovafloxacin, vancomycin, aldosterone, beclometasone, Betamethasone Valerate, reflunomide, hydrocortisone, acetic acid cortisone, acetic acid Desoxycortone, dexamethasone, acetic acid fluohydrocortisone, glucocorticoids, hydrocortisone, methylprednisolone, prednisolone, prednisone, steroid and triamcinolone, antiviral agent, comprises such as Abacavir, acyclovir, acyclovir, Adefovir, amantadine, amprenavir, antiretroviral fixed dosage combines, antiretroviral works in coordination with toughener, Arbidol, Reyataz R, atorvastatincalcuim, brivudine, cidofovir, Combivir, DRV, Delavirdine, Didanosine, how can husky promise, edoxudine, efavirenz, emtricitabine, enfuirtide, Entecavir, entry inhibitor, Famciclovir, Fomivirsen, Fosamprenavir, FOSCARNET, PHOSPHONACETIC, fusion inhibitor, ganciclovir, Jia Dexi, ibacitabine, iodoxuridine, Imiquimod, isoprinosine, Indinavir, inosine, integrase inhibitor, Interferon, rabbit, I type Interferon, rabbit, II type Interferon, rabbit, type iii interferon, lamivudine, rltonavir, loviride, Maraviroc, MK-0518, Moroxydine, viracept see nelfinaivr, nevirapine, Nexavar, nucleoside analog, Oseltamivir, Penciclovir, Peramivir, Pu Kenali, podophyllotoxin, proteinase inhibitor, reverse transcriptase inhibitors, virazole, Rimantadine, ritonavir, Saquinavir, stavudine, tynofovir, tenofovir disoproxil, tipranavir, trifluridine, three associations only, tromantadine, Troyes reaches, valacyclovir, valganciclovir, Wei Kelinuo, vidarabine, the big miaow that draws is fixed, zalcitabine, zanamivir, zidovudine, or its any combination,

(9) another therapeutical agent described is selected from cytotoxic agent, chemotherapeutic or immunosuppressor, and it comprises: 1-boldenone, 1-methyl-nitroso-urea, 5 FU 5 fluorouracil, Ismipur, Ismipur, 6-Tioguanine, Orencia, abraxane, Etretin, aclarubicin, actinium-225 ( ²²⁵ac), actinomycin, adalimumab, adenosine deaminase inhibitors, Afelimomab, VEGF Trap, atropic pearl monoclonal antibody, Ah method's Saite, alitretinoin, alkyl sulfonate esters, alkylating agent, altretamine, ground, Avobenzene west, amino-laevulic acid/amino-laevulic acid methyl esters, aminopterin-induced syndrome, aminopterin-induced syndrome, amrubicin, amsacrine, amsacrine, anagrelide, Kineret, amerantrone class, anthracyclines, anthracyclines, anthracyclines, Antramycin (AMC), antimitotic agent, microbiotic, anti-CD20 antibodies, antifol, antilymphocyte globulin (ALG), antimetabolite, antithymocyte globulin, white arsenic, A Sai pearl monoclonal antibody, asparaginase, fall l-asparagine medicine, astatine-211 ( ²¹¹at), A Li pearl monoclonal antibody, atorolimumab, atrasentan, Avastin ^tM, azacitidine, azathioprine, spit of fland, nitrogen Lars, aziridines, basiliximab, BAYX antibody, Bei Laxipu, Baily monoclonal antibody, Belotecan, bendamustine, cypress for wooden monoclonal antibody, bexarotene, bisantrene, bismuth-213 ( ²¹³bi), bismuth-212 ( ²¹²bi), bleomycin, bleomycin, bleomycin, BLyS antibody, Velcade, busulfan, busulfan, Calcineurin inhibitors, calicheamicin, camptothecine, camptothecin, capecitabine, carboplatin (Paraplatin), carboquone, carminomycin, carmofur, carmustine, carmustine (BSNU), CAT antibody, CD11a antibody, CD147/ fundamental immunity globulin antibody, CD154 antibody, CD18 antibody, CD20 antibody, CD23 antibody, CD3 antibody, CD4 antibody, CD40 antibody, CD62L/L-selects plain antibody, CD80 antibody, CDK inhibitor, cedelizumab, celecoxib, Pegylation match trastuzumab, Chlorambucil, Chlorambucil class, S-Neoral, cis-dichlorodiamine platinum (II) (DDP) cis-platinum, CldAdo, clenoliximab, Clofarex, colchicine, complement component 5 antibody, copper-67 (67Cu), reflunomide, CTLA-4 antibody, CTLA-4 fused protein, cyclophilin inhibitor, endoxan, epithio phosphamide, cytosine arabinoside, cytosine arabinoside, cytochalasin B, cytotoxicity rnase, Dacarbazine, daclizumab, gengshengmeisu, gengshengmeisu (dactinomycin), daunomycin, daunomycin, daunomycin (claiming daunorubicin in the past), Decitabine, De Fulimosi, Omaine, detorubicin, mitobronitol, diethylcarbamazine, dihydrofolate reductase inhibitor, dihydroxyl anthracin diketone, diphtheria toxin, archaeal dna polymerase inhibitor, Docetaxel, Dorlimomab Aritox, to get profit western pearl monoclonal antibody, Dx (Zorubicin), DXL625, according to storehouse pearl monoclonal antibody, efalizumab, second method former times sieve, EGFR antagonist, department of Erie is not, elsamitrucin, Ai Ximo monoclonal antibody, Hemometine, endothelin-receptor antagonists, epipodophyllotoxin class, epirubicin, ebormycine class, Erbitux ^tM, the sharp pearl monoclonal antibody of strategic point, Emcyt, etanercept, ethidium bromide, etoglucid, Etoposide, etoposide phosphate, everolimus, faralimomab, farnesyl tranfering enzyme inhibitor, FKBP inhibitor, floxuridine, fludarabine, Fluracil, virtue trastuzumab, fotemustine, galiximab, gallium-67 (67Ga), Ge Telu monoclonal antibody, Jia Weimo monoclonal antibody, gemcitabine, glucocorticoids, the wooden monoclonal antibody of dagger-axe profit, Shandong former times monoclonal antibody, Gramicidin D, gusperimus, hydrazine class, hydroxyurea, hypomethylation agent, idarubicin, idarubicin, ifosfamide, IL-I antagonist, IL-I receptor antagonist, IL-12, IL-12 antibody, IL-12R antagonist, IL-13 antibody, IL-2, IL-2 inhibitor, IL-2 acceptor/CD25 antibody, IL-6 antibody, imatinib mesylate, immunoglobulin E antibodies, IMP dehydrogenase inhibitor, infliximab, Inolimomab, integrin antibody, interferon antibody, Interferon, rabbit, interleukin-15 antibody, interleukin-6 receptor antibody, interleukin, iodine-125 ( ¹²⁵i), iodine-131 (131I), her monoclonal antibody, irinotecan, ipsapirone, keliximab, La Luotasai, lead-212 ( ²¹²pb), Lai Buli pearl monoclonal antibody, leflunomide, Revlimid, lerdelimumab, formyl tetrahydrofolic acid, LFA-I antibody, lignocaine, lipoxidase inhibitor, lomustine (CCNU), lonidamine, lucanthone, Shandong former times monoclonal antibody, lutetium-177 ( ¹⁷⁷lu), Macrolide, Mannosulfan, Maslimomab, masoprocol, mechlorethamine, melphalan, mepolizumab, purinethol, beautiful for wooden monoclonal antibody, methotrexate, microtubules inhibitor, microtubule stability toughener, mithramycin, mitobronitol, mitoguazone, mitomycin, ametycin, mitotane, mitoxantrone, morolimumab, mTOR inhibitors, Muromondb-CD3, nitrogen mustards, mycophenolic acid, mitotane (O, P'-(DDD)), natalizumab, S 254, nerelimomab, nimustine, nitrogen mustards, nitrosourea, positive dihydroguaiaretic acid, Ao Limosen, auspicious pearl monoclonal antibody difficult to understand, auspicious pearl monoclonal antibody difficult to understand, Odulimomab, method wood monoclonal antibody difficult to understand, Aura handkerchief Buddhist nun, omalizumab, Ao Tasai, Austria's former times pearl monoclonal antibody, oxaliplatin, oxaliplatin, taxol (PTX), pearl monoclonal antibody examined by handkerchief, PDGF antagonist, pegaspargase, pemetrexed, spray department statin, handkerchief trastuzumab, training gram pearl monoclonal antibody, phosphodiesterase inhibitor, phosphorus-32 ( ³²p), pimecrolimus abetimus, pirarubicin, China fir fine jade, platinum class, Plicamycin, Poly ADP-ribose polymerase inhibitor, porfimer sodium, derivatives of porphyrin, PM, PROCAINE HCL, PHARMA GRADE, Procarbazine, Procarbazine, Proprasylyte, proteasome inhibitor, Pseudomonas exotoxin, pseudomonal toxin, purine synthetic inhibitor, tetracycline, pyrimidine synthesis inhibitors, radionuclide, radiotherapy dose, Raltitrexed, ranomustine, Rayleigh pearl monoclonal antibody, retinoid X receptor agonist, class xanthoplane, rhenium-186 ( ¹⁸⁶re), rhenium-188 ( ¹⁸⁸re), ribonucleotide reductase inhibitors, Ricin, Li Naxipu, rovelizumab, Rubitecan, ruplizumab, samarium-153 ( ¹⁵³sm), Satraplatin, scandium-47 ( ⁴⁷sc), SARM, selective estrogen receptor modulators, fill in sharp Seeley, semustine, sexual hormoue antagonist, cedelizumab, sirolimus, steroidal aromatase inhibitor, steroid, streptozocin, U-9889, tacrolimus, talaporfin, his sharp pearl monoclonal antibody, taxanes, PTX class, Ftorafur, telimomab aritox, temoporfin, Temozolomide, CCI-779, CCI-779, for how former times monoclonal antibody, teniposide, for sharp pearl monoclonal antibody, Teriflunomide, for Si Tasai, testolactone, tetracaine, thalidomide, thiophene is for sending Chlorambucil, thio-purine Tioguanine, thiophene is for group, thymidylate synthetase inhibitor, tiazofurine, for pyrrole method Buddhist nun, T-lymphocyte antibody, TNF antagonist, TNF antibody, TNF fused protein, TNF receptor fusion protein matter, TNF-alpha inhibitor, trastuzumab, topoisomerase enzyme inhibitor, topotecan, hold in the palm sharp pearl monoclonal antibody, ET-743, Sibutramine Hydrochloride wood monoclonal antibody, Treosulfan, vitamin A acid, triazene, triaziquone, Tretamine, four nitric acid three platinum, trofosfamide, specific for tumour antigen monoclonal antibody, tyrosine kinase inhibitor, Uramustine, excellent spy gram monoclonal antibody, valrubicin, valrubicin, cut down profit former times monoclonal antibody, VEGF antagonist, vepalimomab, Visudyne, vinealeucoblastine(VLB), vinca alkaloids, vincristine(VCR), vindesine, Vinflunine, vinorelbine, tie up western pearl monoclonal antibody, Vorinostat, yttrium-88 ( ⁸⁸y), Yttrium-90 ( ⁹⁰y), prick wooden monoclonal antibody, zileuton, draw wooden monoclonal antibody, zolimomab aritox, zorubicin, Zuo Tamosi or its any combination together, and/or

(10) another promoting agent described is comprise one or more agonists of the following factor, antagonist or conditioning agent: TNF-α, IL-2, IL-4, IL-6, IL-10, IL-12, IL-13, IL-18, element adjusted by WN-γ, BAFF, CXCL13, IP-10, VEGF, EPO, EGF, HRG, pHGF (HGF), iron, or any combination of (1) to (10).