RU2851121C1 - New bifunctional multispecific antagonists capable of inhibiting multiple ligands of the tgf-beta family and their applications - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to a selected molecule of a multispecific antagonist containing a first antigen-binding molecule that specifically binds to activin or the activin receptor, and a second antigen-binding molecule that specifically binds to the TGF-β receptor, wherein said multispecific antagonist molecule simultaneously neutralises the activin signalling pathway and the TGF-β signalling pathway, wherein the molecule of said multispecific antagonist contains an amino acid sequence selected from the group consisting of the amino acid sequences represented in SEQ ID NO: 54, 55, 60 and 61.

EFFECT: pharmaceutical composition containing such a multispecific antagonist is proposed, which is intended for the treatment of a painful condition involving overexpression of TGF-β and activin, such as cancer.

6 cl, 15 dwg, 11 tbl, 6 ex

Description

Related patent applications

This patent application claims the benefit of U.S. Provisional Patent Application No. 63/113,920, filed November 15, 2020, and U.S. Provisional Patent Application No. 63/109,814, filed November 4, 2020, each of which is incorporated herein by reference.

Prior art

The transforming growth factor beta (TGF-β) superfamily consists of TGF-β proteins, activins, growth differentiation factors (GDFs), bone morphogenetic proteins (BMPs), glial cell-derived neurotrophic factors (GDNFs), inhibins, lymph node-derived factors, Lefty (left and right differentiation factor), and Müllerian inhibitory substance (MIS). TGF-β superfamily ligands form dimers that bind to heterodimeric receptor complexes composed of type I and type II receptor subunits with serine/threonine kinase domains. Upon ligand binding, the type II receptor phosphorylates and activates the type I receptor, initiating a Smad-dependent signaling cascade that induces or suppresses transcriptional activity.

TGF-β (TGF-β1, TGF-β2, and TGF-β3) mediate the Smad2/3 signaling pathway by binding to and activating the high-affinity cell surface receptors TGFβRII and TGFβRIIB. TGF-β plays a critical role in regulating a wide range of biological activities, including immune function, cell proliferation and differentiation, fibrogenesis, epithelial-mesenchymal transition, hematopoiesis, myogenesis, and bone remodeling. Elevated TGF-β levels and subsequently enhanced Smad2/3 signaling are implicated in the pathogenesis and progression of numerous disease states, including cancer, fibrosis, anemia, bone metastasis, bone loss, pain, muscle wasting, insulin resistance, chronic kidney disease, liver disease, and cardiovascular disease.

Additionally, as a subgroup of the TGF-β superfamily, activins and related proteins (including activin A, activin B, activin AB, GDF-8, and GDF-11) also mediate the Smad2/3 signaling pathway by binding to and activating their high-affinity cell surface receptors, ActRIIA and ActRIIB. Activin and related proteins regulate a wide range of biological activities, including immune function, cellular differentiation, myogenesis, fibrogenesis, bone remodeling, hematopoiesis, and reproductive physiology. Follistatin (FST), a secreted glycoprotein, binds to activins and activin-related ligands to negatively control their signaling activities. Overexpression of activins and related ligands and increased Smad2/3 signaling are implicated in the pathogenesis and progression of many severe conditions such as cancer, fibrosis, anemia, bone metastasis, bone fragility, fracture, muscle atrophy, wasting, pulmonary hypertension, pain, insulin resistance, chronic renal failure, liver disease, myocardial infarction, and heart failure.

The TGF-β/Activin-Smad2/3 signaling pathway plays a central role in the pathogenesis and progression of fibrosis. A key mechanism underlying the pathogenesis and progression of fibrosis is elevated TGF-β/Activin/Smad2/3 signaling, which leads to fibroblast proliferation and activation and, consequently, overexpression of extracellular matrix components such as COL1A1, COL1A2, COL3A1, COL5A2, COL6A1, and COL6A3 in diseased tissue. Evidence suggests that TGF-β and activin are upregulated during fibrosis. When elevated, TGF-β or activin can induce fibroblast activation, leading to fibrosis. Since both TGF-β and activin levels are increased, it is important to inhibit not only TGF-β but also activin to more effectively attenuate fibrosis.

The TGF-β/activin Smad2/3 signaling pathways also play an important role in cancer pathogenesis and progression, in part due to their observed effects on tumor immune surveillance. TGF-β and Activin are highly expressed in various cancers. Overexpression of TGF-β and activin in the cancer microenvironment leads to activation of the Smad2/3 signaling pathway in regulatory T cells (Tregs), leading to their proliferation and activation. In turn, increased Tregs activity causes inhibition of CD8+ cytotoxic T cells, thereby reducing the ability of CD8+ cytotoxic T cells to detect and kill cancer cells. Currently, many clinical trials in immuno-oncology are focused on TGF-β inhibitors and T-cell immune checkpoint inhibitors (such as anti-PD-1 antibodies and anti-CTLA4 antibodies). However, it is important to note that TGF-β and activin are overexpressed in various cancers, and like increased TGF-β, increased activin, or activin-related ligands, they can also suppress cancer immune surveillance by activating Tregs. Thus, the concomitant inhibition of increased TGF-β levels and increased activin levels may more effectively attenuate Tregs and thus more effectively restore immune surveillance in cancer. Thus, a multispecific polypeptide antagonist capable of neutralizing not only increased TGF-β (i.e., TGF-β 1, TGF-β2, and TGF-β3) but also increased activins (i.e., activin A, activin B, and activin AB) in cancer may result in superior efficacy and improved response rates to cancer treatment when co-administered with an immune checkpoint inhibitor.

Description of the invention

In one aspect of the present invention, novel polypeptide-based multispecific antagonist molecules are provided that are specifically designed to simultaneously neutralize the activin signaling pathway and the TGF-β signaling pathway in an effective manner. In various embodiments, the multispecific antagonist molecule is designed as depicted in FIGS. 1-4.

In various embodiments, the multispecific antagonist is a multispecific antagonist molecule comprising a first antigen-binding molecule that specifically binds to an activin ligand or activin-related ligand (an "activin-binding polypeptide") and a second antigen-binding molecule that specifically binds to a TGF-β ligand (a "TGF-β-binding polypeptide"). In various embodiments, the "activin-binding polypeptide" is selected from any polypeptide that is capable of binding to activin (i.e., activin A, activin B, or activin AB) and/or an activin-related ligand (i.e., GDF8 or GDF11), which includes, but is not limited to, an anti-activin antibody (including an anti-activin A antibody and an anti-activin B antibody), a fragment of an anti-activin antibody, the extracellular domains (ECDs) of the wild-type activin receptor type 2 A (ActRIIA) or the wild-type activin receptor type 2 B (ActRIIB), modified extracellular domains of ActRIIA and ActRIIB, native wild-type and modified activin-binding proteins such as follistatin, follistatin-like protein and propeptide, and a phage-display-derived polypeptide targeted against activin or an activin-related ligand, and the "TGF-β-binding polypeptide" is selected from the group consisting of anti-TGF-β antibodies, anti-TGF-β antibody fragments, extracellular domains (ECDs) of wild-type TGF-β receptors type 2 (including TGFβRIIA and TGFβRIIB), modified extracellular domains of TGFβRIIA and TGFβRIIB, and a phage-display-derived antagonist polypeptide targeted against a TGF-β ligand.

In various embodiments, the multispecific antagonist molecule comprises an isolated antibody or antigen-binding fragment thereof that specifically binds to activin or an activin-related ligand, and an isolated antibody or antigen-binding fragment thereof that specifically binds to a TGF-β ligand. In various embodiments, the isolated antibody or antigen-binding fragment thereof is selected from the group consisting of monoclonal Abs (mAbs), polyclonal Abs, Ab fragments (e.g., Fab, Fab', F(ab')2, Fv, Fc, etc.), chimeric Abs, mini-Abs or domain Abs (dAbs), dual-specific Abs, bispecific Abs, heteroconjugate Abs, single-chain Abs (SCAs), single-chain variable region fragments (ScFv), humanized Abs, fully human Abs, and any other modified configuration of an immunoglobulin (Ig) molecule that contains a site recognizing an antigen with the required specificity. In various embodiments, the multispecific molecule comprises an isolated antibody or antigen-binding fragment thereof selected from the group consisting of a fully human, a humanized, and a chimeric antibody.

In various embodiments, the first antigen-binding molecule specifically binds to activin or an activin-related ligand comprising the amino acid sequence presented in SEQ ID NO: 1. In various embodiments, the first antigen-binding molecule specifically binds to activin or an activin-related ligand comprising the amino acid sequence presented in SEQ ID NO: 2. In various embodiments, the first antigen-binding molecule specifically binds to activin or an activin-related ligand comprising the amino acid sequence presented in SEQ ID NO: 3. In various embodiments, the first antigen-binding molecule specifically binds to activin or an activin-related ligand comprising the amino acid sequence presented in SEQ ID NO: 4. In various embodiments, the first antigen-binding molecule specifically binds to activin or an activin-related ligand comprising the amino acid sequence presented in SEQ ID NO: 5. In various embodiments, the first the antigen-binding molecule specifically binds to activin or an activin-related ligand comprising the amino acid sequence presented in SEQ ID NO: 6. In various embodiments, the first antigen-binding molecule specifically binds to activin or an activin-related ligand comprising the amino acid sequence presented in SEQ ID NO: 7. In various embodiments, the first antigen-binding molecule specifically binds to activin or an activin-related ligand comprising the amino acid sequence presented in SEQ ID NO: 8. In various embodiments, the first antigen-binding molecule specifically binds to activin or an activin-related ligand comprising the amino acid sequence presented in SEQ ID NO: 9.

In various embodiments, the first antigen-binding molecule that specifically binds to activin or an activin-related ligand is an isolated antibody selected from the group consisting of an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 10; an antibody comprising the light chain amino acid sequence shown in SEQ ID NO: 11; an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 10 and the light chain amino acid sequence shown in SEQ ID NO: 11; an antibody comprising the heavy chain variable region amino acid sequence shown in SEQ ID NO: 12; an antibody comprising the light chain variable region amino acid sequence shown in SEQ ID NO: 13; and an antibody comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 12 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 13.

In various embodiments, the first antigen-binding molecule that specifically binds to activin or an activin-related ligand is an isolated antibody selected from the group consisting of an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 14; an antibody comprising the light chain amino acid sequence shown in SEQ ID NO: 15; an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 14 and the light chain amino acid sequence shown in SEQ ID NO: 15; an antibody comprising the heavy chain variable region amino acid sequence shown in SEQ ID NO: 16; an antibody comprising the light chain variable region amino acid sequence shown in SEQ ID NO: 17; and an antibody comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 16 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 17.

In various embodiments, the second antigen-binding molecule specifically binds to a TGF-β ligand comprising the amino acid sequence presented in SEQ ID NO: 18. In various embodiments, the second antigen-binding molecule specifically binds to a TGF-β ligand comprising the amino acid sequence presented in SEQ ID NO: 19. In various embodiments, the second antigen-binding molecule specifically binds to a TGF-β ligand comprising the amino acid sequence presented in SEQ ID NO: 20. In various embodiments, the second antigen-binding molecule specifically binds to a TGF-β ligand comprising the amino acid sequence presented in SEQ ID NO: 21.

In various embodiments, the second antigen-binding molecule that specifically binds to a TGF-β ligand is an isolated antibody selected from the group consisting of an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 22; an antibody comprising the light chain amino acid sequence shown in SEQ ID NO: 23; an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 22 and the light chain amino acid sequence shown in SEQ ID NO: 23; an antibody comprising the heavy chain variable region amino acid sequence shown in SEQ ID NO: 24; an antibody comprising the light chain variable region amino acid sequence shown in SEQ ID NO: 25; and an antibody comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 24 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 25.

In various embodiments, the multispecific antagonist is a polypeptide molecule comprising a first antigen-binding molecule that specifically binds to an activin ligand and a second antigen-binding molecule that specifically binds to a TGF-β ligand, wherein the activin ligand-binding molecule is selected from the group of polypeptides comprising the amino acid sequence presented in SEQ ID NOs: 1-17, and the TGF-β ligand-binding molecule is selected from the group of polypeptides comprising the amino acid sequence presented in SEQ ID NOs: 18-25.

In various embodiments, the multispecific antagonist is a polypeptide molecule comprising a first antigen-binding molecule that specifically binds to an activin ligand and a second antigen-binding molecule that specifically binds to a TGF-β ligand, wherein the multispecific molecule is selected from the group consisting of: a multispecific molecule comprising a heavy chain selected from a heavy chain comprising an amino acid sequence presented in SEQ ID NOs: 72-84, and a light chain selected from a light chain comprising an amino acid sequence presented in SEQ ID NOs: 11, 13, 15, 17, 19, 21, 23, and 25.

In various embodiments, the multispecific antagonist is a multispecific antagonist molecule comprising a first antigen-binding molecule that specifically binds to an activin ligand and a second antigen-binding molecule that specifically binds to a TGF-β ligand, wherein the multispecific molecule is selected from the group consisting of a multispecific polypeptide molecule comprising the amino acid sequence presented in SEQ ID NO: 54-71.

In another aspect, the present invention provides novel bifunctional multispecific polypeptide-based antagonist molecules specifically designed to simultaneously inhibit TGF-β, activin, and a T cell immune checkpoint (including PD1, PDL1, or CTL4). In various embodiments, the bifunctional multispecific antagonist is a bifunctional multispecific molecule comprising a first antigen-binding molecule that specifically binds to an activin ligand or an activin-related ligand (an "activin-binding polypeptide"), a second antigen-binding molecule that specifically binds to a TGF-β ligand (a "TGF-β-binding polypeptide"), and a third antigen-binding molecule that specifically binds to a PD-1 ligand, a PD-L1 ligand, or a CTLA-4 ligand (a "PD-1/PD-L1/CTLA-4-binding polypeptide"). In various embodiments, the bifunctional multispecific antagonist molecule is designed as depicted in FIGS. 5-6.

In various embodiments, the third antigen-binding molecule that specifically binds to a PD-1 ligand is an isolated antibody selected from the group consisting of an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 26; an antibody comprising the light chain amino acid sequence shown in SEQ ID NO: 27; an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 26 and the light chain amino acid sequence shown in SEQ ID NO: 27; an antibody comprising the heavy chain variable region amino acid sequence shown in SEQ ID NO: 28; an antibody comprising the light chain variable region amino acid sequence shown in SEQ ID NO: 29; and an antibody comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 28 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 29.

In various embodiments, the third antigen-binding molecule that specifically binds to a PD-1 ligand is an isolated antibody selected from the group consisting of an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 30; an antibody comprising the light chain amino acid sequence shown in SEQ ID NO: 31; an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 30 and the light chain amino acid sequence shown in SEQ ID NO: 31; an antibody comprising the heavy chain variable region amino acid sequence shown in SEQ ID NO: 32; an antibody comprising the light chain variable region amino acid sequence shown in SEQ ID NO: 33; and an antibody comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 32 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 33.

In various embodiments, the third antigen-binding molecule that specifically binds to a PD-1 ligand is an isolated antibody selected from the group consisting of an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 38; an antibody comprising the light chain amino acid sequence shown in SEQ ID NO: 39; an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 38 and the light chain amino acid sequence shown in SEQ ID NO: 39; an antibody comprising the heavy chain variable region amino acid sequence shown in SEQ ID NO: 40; an antibody comprising the light chain variable region amino acid sequence shown in SEQ ID NO: 41; and an antibody comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 40 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 41.

In various embodiments, the third antigen-binding molecule that specifically binds to a PD-1 ligand is an isolated antibody selected from the group consisting of an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 42; an antibody comprising the light chain amino acid sequence shown in SEQ ID NO: 43; an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 42 and the light chain amino acid sequence shown in SEQ ID NO: 43; an antibody comprising the heavy chain variable region amino acid sequence shown in SEQ ID NO: 44; an antibody comprising the light chain variable region amino acid sequence shown in SEQ ID NO: 45; and an antibody comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 44 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 45.

In various embodiments, the third antigen-binding molecule that specifically binds a CTLA-4 ligand is an isolated antibody selected from the group consisting of an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 34; an antibody comprising the light chain amino acid sequence shown in SEQ ID NO: 35; an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 34 and the light chain amino acid sequence shown in SEQ ID NO: 35; an antibody comprising the heavy chain variable region amino acid sequence shown in SEQ ID NO: 36; an antibody comprising the light chain variable region amino acid sequence shown in SEQ ID NO: 37; and an antibody comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 36 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 37.

In various embodiments, the third antigen-binding molecule that specifically binds to a PD-L1 ligand is an isolated antibody selected from the group consisting of an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 46; an antibody comprising the light chain amino acid sequence shown in SEQ ID NO: 47; an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 46 and the light chain amino acid sequence shown in SEQ ID NO: 47; an antibody comprising the heavy chain variable region amino acid sequence shown in SEQ ID NO: 48; an antibody comprising the light chain variable region amino acid sequence shown in SEQ ID NO: 49; and an antibody comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 48 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 49.

In various embodiments, the third antigen-binding molecule that specifically binds to a PD-L1 ligand is an isolated antibody selected from the group consisting of an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 50; an antibody comprising the light chain amino acid sequence shown in SEQ ID NO: 51; an antibody comprising the heavy chain amino acid sequence shown in SEQ ID NO: 50 and the light chain amino acid sequence shown in SEQ ID NO: 51; an antibody comprising the heavy chain variable region amino acid sequence shown in SEQ ID NO: 52; an antibody comprising the light chain variable region amino acid sequence shown in SEQ ID NO: 53; and an antibody comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 52 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 53.

In various embodiments, the bifunctional multispecific antagonist is a bifunctional multispecific molecule comprising a first antigen-binding polypeptide that specifically binds to an activin ligand, and a second antigen-binding polypeptide that specifically binds to a TGF-β ligand, and a third antigen-binding polypeptide that specifically binds to a PD-1 ligand, a PD-L1 ligand, or a CTLA-4 ligand, wherein the bifunctional multispecific molecule is selected from the group consisting of: a bifunctional multispecific molecule comprising a heavy chain selected from a heavy chain comprising the amino acid sequence presented in SEQ ID NOs: 85-120, and a light chain selected from a light chain comprising the amino acid sequence presented in SEQ ID NOs: 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51 and 53. In various embodiments, the bifunctional multispecific antagonist molecule is a bifunctional multispecific antagonist molecule comprising a heavy chain selected from a heavy chain comprising an amino acid sequence presented in SEQ ID NO: 85-102 and a light chain comprising an amino acid sequence presented in SEQ ID NO: 27. In various embodiments, the bifunctional multispecific antagonist molecule is a bifunctional multispecific antagonist molecule comprising a heavy chain selected from a heavy chain comprising an amino acid sequence presented in SEQ ID NO: 103-120 and a light chain comprising an amino acid sequence presented in SEQ ID NO: 35.

In another aspect of the present description, a pharmaceutical composition is provided comprising isolated molecules of multispecific or bifunctional multispecific antagonists in admixture with a pharmaceutically acceptable carrier.

In another aspect of the present disclosure, a method for treating or preventing various complex disease states, the pathogenesis of which involves parallel activation of the activin signaling pathway and the TGF-β signaling pathway, is provided.

In various embodiments, the novel multispecific antagonists or bifunctional multispecific antagonist molecules of the present invention may have broad applications in the treatment of a variety of disorders, including, but not limited to, the following conditions: blood disorders: ineffective erythropoiesis, anemia, pancytopenia, myelodysplastic syndromes; fibrotic diseases: non-alcoholic steatohepatitis/NASH, liver fibrosis, pulmonary fibrosis, renal fibrosis, polycystic kidney disease, cardiac fibrosis, muscle fibrosis, myelofibrosis, cutaneous fibrosis, and ocular fibrosis; hematological malignancies: multiple myeloma, leukemia, and lymphoma; Solid cancer: melanoma, multiple myeloma, lung cancer, pancreatic cancer, colorectal cancer, liver cancer, stomach cancer, kidney cancer, bladder cancer, head and neck cancer, thyroid cancer, breast cancer, ovarian cancer, endometrial cancer, testicular cancer, prostate cancer, brain cancer and sarcoma; Metastasis: bone metastasis, lung metastasis, liver metastasis, brain metastasis; Cancer treatment in combination with checkpoint inhibitors such as anti-PD-1, anti-PDL1 and anti-CTL4 antibodies; Neuromuscular diseases: muscular dystrophy, spinal muscular atrophy, spinal cord injury, stroke; Pain: nociceptive pain, neuropathic pain; Wasting syndromes: sarcopenia, cancer wasting, cardiac wasting, renal wasting, rheumatoid wasting and anorexia; Bone disorders: bone metastasis, skeletal phenomena in cancer, bone fragility, fracture, osteogenesis imperfection, and osteoporosis; Cardiovascular diseases: pulmonary hypertension, myocardial infarction, heart failure; Metabolic disorders: insulin resistance, diabetic nephropathy, chronic renal failure; Inflammatory diseases: rheumatoid arthritis, inflammatory bowel disease; Infections: SARS-CoV (severe acute respiratory syndrome coronavirus), cytokine storm syndrome, sepsis; and Trauma: burn injury.

In another aspect of the disclosure, uses of bifunctional antagonist molecules or bifunctional multispecific antagonist molecules for the preparation of a medicament for the treatment of any disorder or condition described herein are provided.

In another aspect, the present disclosure provides isolated nucleic acid molecules comprising a polynucleotide encoding a multispecific antagonist molecule or a bifunctional multispecific antagonist molecule as described herein. In various embodiments, the isolated nucleic acid molecules comprise the polynucleotides described herein and further comprise a polynucleotide encoding at least one heterologous protein described herein. In various embodiments, the nucleic acid molecules further comprise polynucleotides encoding linkers or hinge linkers as described herein.

Another aspect of the present disclosure provides vectors comprising the nucleic acids described herein. In various embodiments, the vector is an expression vector. Another aspect of the present disclosure provides isolated cells comprising nucleic acids according to the disclosure. In various embodiments, the cell is a host cell comprising an expression vector according to the disclosure. Another aspect provides methods for producing multispecific antagonist molecules or bifunctional multispecific antagonist molecules by culturing host cells under conditions conducive to the expression of proteins or polypeptides.

In another aspect, a method is provided for producing a multispecific antagonist molecule or a bifunctional multispecific antagonist molecule comprising a first antigen-binding molecule that specifically binds to activin or an activin-related ligand, such as GDF8 or GDF11, and a second antigen-binding molecule that specifically binds to TGF-β, as described herein, comprising the steps of a) transforming a host cell with vectors containing polynucleotides encoding said multispecific antagonist molecule or bifunctional multispecific antagonist molecule, b) growing the host cell in accordance with conditions suitable for expressing the multispecific antagonist molecule or bifunctional multispecific antagonist molecule, and c) isolating the bifunctional antagonist molecule from the culture. The invention also encompasses a multispecific antagonist molecule or a bifunctional multispecific antagonist molecule obtainable by the method according to the invention.

Brief description of graphic materials

FIG. 1 represents a typical multispecific antagonist molecule according to the present invention. The "activin-binding polypeptide" illustrated in this diagram refers to any polypeptide that is capable of binding to activin (i.e., activin A, activin B, or activin AB) and/or an activin-related ligand (i.e., GDF8 or GDF11), which includes, but is not limited to, 1) an anti-activin antibody (including an anti-activin A antibody and an anti-activin B antibody), a fragment of an anti-activin antibody, the extracellular domains (ECDs) of the wild-type activin receptor type 2A (ActRIIA) or activin receptor type 2B (ActRIIB), the extracellular domains of modified ActRIIA and ActRIIB, native wild-type and modified activin-binding proteins such as follistatin, follistatin-like protein, and propeptide, and a phage-display-derived polypeptide capable of binding and blocking activin and/or an activin-related ligand. "TGF-β binding polypeptide" illustrated in this diagram refers to any polypeptide that is capable of binding to TGF-β (i.e., TGF-β1, TGF-β2, or TGF-β3), which includes, but is not limited to, an anti-TGF-β antibody, an anti-TGF-β antibody fragment, the extracellular domains (ECDs) of wild-type TGF-β type-2 receptors (including TGFβRIIA and TGFβRIIB), the extracellular domains of modified TGFβRIIA and TGFβRIIB, and phage display-derived antagonist polypeptides that are capable of binding and neutralizing TGF-β. "Linker" illustrated in this diagram refers to various methods of fusing different polypeptide fusion partners to produce bispecific and multispecific molecules, which includes, but is not limited to, the use of any peptide linker or chemical linker.

FIG. 2 shows two bifunctional antagonist molecules of the present invention in different configurations (FIGS. 2A and 2B), wherein the activin-binding polypeptide is the ECD of the activin receptor and the TGF-β-binding polypeptide is the ECD of the TGF-β receptor linked via an Fc domain.

FIG. 3A and 3B represent two exemplary bifunctional antagonist molecules according to the present invention, wherein: (A) the activin-binding polypeptide is an anti-activin antibody and the TGF-β-binding polypeptide is the ECD of the TGF-β receptor linked via a linker to the CH3 of the heavy chain of an anti-activin antibody; or (B) the TGF-β-binding polypeptide is an anti-TGF-β antibody and the activin-binding polypeptide is the ECD of the activin receptor linked via a linker to the CH3 of the heavy chain of an anti-TGF-β antibody. In alternative embodiments, the TGF-β receptor ECD (or the TNF receptor ECD) is linked to the anti-activin antibody (or the anti-TGF-β antibody) via a linker in the heavy chain variable region (VH) of the antibody. In alternative embodiments, the TGF-β receptor ECD (or the activin receptor ECD) is linked to the anti-activin antibody (or anti-TGF-β antibody) via a linker in the variable region of the light chain (VL) of the antibody. In alternative embodiments, the TGF-β receptor ECD (or the TNF receptor ECD) is linked to the anti-activin antibody (or anti-TGF-β antibody) via a linker at an internal site, rather than at the CH3, VL, or VH sites of the antibody's heavy chain.

FIG. 4 shows a bifunctional antagonist molecule according to the present invention in the form of a bispecific antibody, wherein the activin-binding polypeptide is represented by the variable regions (V _L and V _H ) of an anti-activin A antibody and the TGF-β-binding polypeptide is represented by the variable regions (V _L and V _H ) of an anti-TGF-β antibody. The bispecific antibody shown in FIG. 4 is only an example, since bispecific antibodies comprising the variable regions of an anti-TGF-β antibody and anti-activin antibodies can be designed in various configurations that differ from the example shown in FIG. 4.

FIG. 5 depicts an exemplary novel bifunctional multispecific antagonist molecule according to the present invention capable of inhibiting the T cell immune checkpoint, activin, and TGF-β by fusion between an immune checkpoint inhibitor polypeptide (e.g., the heavy chain of an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody) (fusion partner A), an activin-binding polypeptide (fusion partner B), and a TGFβ-binding polypeptide (fusion partner C). The "immune checkpoint inhibitor polypeptide" illustrated in this diagram refers to any antagonist polypeptides capable of binding to and inhibiting PD-1, PD-L1, or CTLA-4, which includes, but is not limited to, anti-PD-1, anti-PDL1, and anti-CTLA4 antibodies or antibody fragments and PD-1 receptor polypeptide mimics. The "activin-binding polypeptide" illustrated in this diagram refers to any polypeptide that is capable of binding to activin (i.e., activin A, activin B, or activin AB) and/or an activin-related ligand (i.e., GDF8 or GDF11), which includes, but is not limited to, 1) an anti-activin antibody (including an anti-activin A antibody and an anti-activin B antibody), a fragment of an anti-activin antibody, the extracellular domains (ECDs) of the wild-type activin receptor type 2 A (ActRIIA) or activin receptor type 2 B (ActRIIB), the extracellular domains of modified ActRIIA and ActRIIB, native wild-type and modified activin-binding proteins such as follistatin, follistatin-like protein, and propeptide, and a phage-display-derived polypeptide capable of binding and blocking activin and/or an activin-related ligand. "TGFβ-binding polypeptide" as illustrated in the diagram refers to any polypeptide that is capable of binding to TGFβ (i.e., TGFβ1, TGFβ2, or TGFβ3), which includes, but is not limited to, an anti-activin antibody, an anti-TGFβ antibody fragment, extracellular domains (ECDs) of wild-type TGFβ receptors type 2 (including TGFβRIIA and TGFβRIIB), extracellular domains of modified TGFβRIIA and TGFβRIIB, and phage-display-derived antagonist polypeptides that are capable of binding and neutralizing TGFβ. "Linker" as illustrated in the diagram refers to various methods of fusing different polypeptide fusion partners to form bispecific and multispecific molecules, which includes, but is not limited to, the use of any peptide linker or chemical linker.

FIG. 6A and 6B represent two exemplary bifunctional antagonist molecules according to the present invention wherein: (A) the immune checkpoint inhibitor polypeptide is an anti-PD-1 antibody, the activin-binding polypeptide is the ECD of the activin receptor linked via a linker to the CH3 of the heavy chain of the anti-PD-1 antibody, and the TGF-β-binding polypeptide is the ECD of the TGF-β receptor linked via a linker to the ECD of the activin receptor; or (B) the immune checkpoint inhibitor polypeptide is an anti-CTLA-4 antibody, the activin-binding polypeptide is the ECD of the activin receptor linked via a linker to the CH3 of the heavy chain of the anti-CTLA-4 antibody, and the TGF-binding polypeptide is the ECD of the TGF-β receptor linked via a linker to the ECD of the activin receptor.

FIG. 7 presents line graphs demonstrating that multispecific antagonist molecules A115, A116, A117, and A118 effectively neutralize activin A and activin B in cell-based assays. IC ₅₀ values were calculated and plotted using Prism software (GraphPad Software).

FIG. 8 presents line graphs demonstrating that the multispecific antagonist molecules A115, A116, A117, and A118 effectively neutralize TGF-β1 and TGF-β3 in cellular assays. IC ₅₀ values were calculated and plotted using Prism software (GraphPad Software).

FIG. 9 presents line graphs demonstrating that the multispecific antagonist molecules A115, A116, A117, and A118 effectively neutralize GDF-8 and GDF-11 in cell-based assays. IC ₅₀ values were calculated and plotted using Prism software (GraphPad Software).

FIGS. 10-12 show the inhibitory effects of the multispecific antagonist A116 on cell migration and the antagonist A549 on colony formation in human non-small cell lung cancer cell cultures.

FIG. 13 shows histological images of H&E-stained lung sections and Ashcroft scores of lung sections in a mouse model of bleomycin-induced pulmonary fibrosis. The data indicate that dual inhibition of TGF-β and activin with A116 significantly reduced bleomycin-induced pulmonary fibrosis, whereas the activin inhibitor ActRIIA-Fc or the TGF-β blocker TGFRII-Fc moderately inhibited bleomycin-induced pulmonary fibrosis. Ashcroft score analysis revealed that A116 treatment resulted in a greater reduction in pulmonary fibrosis compared to ActRIIA-Fc treatment or TGFRII-Fc treatment.

FIG. 14 shows histological images of Masson trichrome-stained lung sections and a bar graph of the quantitative analysis of collagen deposition areas in lung sections from mice suffering from bleomycin-induced pulmonary fibrosis. These data demonstrate that A116 reduced collagen deposition more effectively than ActRIIA-Fc or TGFRII-Fc in the bleomycin-induced pulmonary fibrosis model.

FIG. 15 shows histological patterns of immunostaining for α-SMA (α-smooth muscle actin), a marker of fibrosis, in lung sections from control and bleomycin-treated mice. Lung sections were immunostained with an anti-α-SMA primary antibody conjugated to an HRP-labeled secondary antibody. The results indicate that A116 was more effective than ActRIIA-Fc or TGFRII-Fc in preventing α-SMA induction in the lungs of bleomycin-treated mice.

Methods for implementing the description of the invention

Definitions

The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. In various embodiments, "peptides," "polypeptides," and "proteins" are chains of amino acids whose alpha carbon atoms are linked via peptide bonds. Thus, the terminal amino acid at one end of the chain (the amino terminus) has a free amino group, while the terminal amino acid at the other end of the chain (the carboxyl terminus) has a free carboxyl group. As used herein, the term "amino terminus" (abbreviated N-terminus) refers to the free α-amino group in the amino acid at the amino terminus of the peptide or to the α-amino group (the imino group involved in peptide bond formation) of the amino acid elsewhere within the peptide. Similarly, the term "carboxyl terminus" refers to a free carboxyl group at the carboxyl terminus of a peptide or to a carboxyl group of an amino acid elsewhere within the peptide. Peptides also include essentially any polyamino acid, including, but not limited to, peptide mimetics, such as amino acids linked via an ether bond, as opposed to an amide bond.

Polypeptides according to this description of the invention include polypeptides that have been modified in any way and for any reason, for example, to: (1) reduce susceptibility to proteolysis, (2) reduce susceptibility to oxidation, (3) change binding affinity for forming protein complexes, (4) change binding affinities, and (5) impart or modify other physicochemical or functional properties.

As used herein, an amino acid "substitution" refers to the replacement of a single amino acid at a specific position in the sequence of a parent polypeptide with a different amino acid. Amino acid substitutions can be made using genetic or chemical methods well known in the art. For example, one or more amino acid substitutions (e.g., conservative amino acid substitutions) can be made in a native sequence (e.g., in a portion of a polypeptide outside the domain(s) that form intermolecular contacts). The term "conservative amino acid substitution" refers to the replacement of an amino acid in a polypeptide with a functionally similar amino acid. Each of the following six groups contains amino acids that are conservative substitutions for one another:

1) Alanine (A), series (S) and threonine (T)

2) Aspartic acid (D) and glutamic acid (E)

3) Asparagine (N) and glutamine (Q)

4) Arginine (R) and lysine (K)

5) Isoleucine (I), leucine (L), methionine (M) and valine (V)

6) Phenylalanine (F), tyrosine (Y) and tryptophan (W)

The term "non-conservative amino acid substitution" refers to the substitution of a member of one of these classes for a member of another class. When making such changes according to the various embodiments, the hydrophobicity index of the amino acids should be taken into account. A hydrophobicity index has been assigned to each amino acid based on its hydrophobicity and charge properties. The indices are: isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine/cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine (-0.4); threonine (-0.7); serine (-0.8); tryptophan (-0.9); tyrosine (-1.3); proline (-1.6); histidine (-3.2); glutamate (-3.5); glutamine (-3.5); aspartate (3.5); asparagine (-3.5); lysine (-3.9) and arginine (-4.5).

The importance of the hydrophobicity index of amino acids in imparting interactive biological function to a protein is understood in the art (see, e.g., Kyte et al., 1982, J. Mol. Biol. 157:105-131). It is known that certain amino acids can be substituted with other amino acids having a similar hydrophobicity index or value, and such amino acids will retain similar biological activity. When making changes based on the hydrophobicity index, various embodiments include substitutions of amino acids whose hydrophobicity indices are within ±2. Various embodiments include substitutions of amino acids whose hydrophobicity indices are within ±1, and various embodiments include substitutions of amino acids whose hydrophobicity indices are within ±0.5.

It is also understood in the art that substitution of such amino acids can be effectively accomplished based on hydrophilicity, particularly when the resulting biologically functional protein or peptide is intended for use in immunological embodiments, as disclosed herein. In various embodiments, the highest local average hydrophilicity of a protein, determined by the hydrophilicity of adjacent amino acids, correlates with its immunogenicity and antigenicity, i.e., with the biological property of that protein.

The following hydrophilicity indices are assigned to the following amino acid residues: arginine (+3.0); lysine (+3.0); aspartate (+3.0 ± 1); glutamate (+3.0 ± 1); serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); threonine (-0.4); proline (-0.5 ± 1); alanine (-0.5); histidine (-0.5); cysteine (-1.0); methionine (-1.3); valine (-1.5); leucine (-1.8); isoleucine (-1.8); tyrosine (-2.3); phenylalanine (-2.5) and tryptophan (-3.4). When making changes based on similar hydrophilicity values, in various embodiments, amino acids with hydrophilicity indices within ±2 are substituted, in various embodiments, amino acids with hydrophilicity indices within ±1 are substituted, and in various embodiments, amino acids with hydrophilicity indices within ±0.5 are substituted.

Examples of amino acid substitutions are presented in Table 1.

One skilled in the art will be able to identify suitable variants of the polypeptides presented herein using well-known methods. In various embodiments, one skilled in the art can identify suitable regions of the molecule that can be altered without disrupting activity by targeting regions that are not believed to be essential for activity. In other embodiments, one skilled in the art can identify residues and fragments of the molecule that are conserved among similar polypeptides. In other embodiments, even regions that may be important for biological activity or structure can undergo conservative amino acid substitutions without disrupting biological activity or adversely affecting the polypeptide structure.

Furthermore, one skilled in the art can consult structure-function studies that identify residues in similar polypeptides that are important for activity or structure. Based on such comparisons, one can predict the importance of amino acid residues in a polypeptide that correspond to amino acid residues important for activity or structure in similar polypeptides. One skilled in the art can select chemically similar amino acid substitutions for such predicted important amino acid residues.

A person skilled in the art can also analyze the three-dimensional structure and amino acid sequence relative to that structure in similar polypeptides. Taking this information into account, a person skilled in the art can predict the alignment of the amino acid residues of a polypeptide relative to its three-dimensional structure. In various embodiments, a person skilled in the art may decide not to make radical changes to amino acid residues that are expected to be on the surface of the polypeptide, since such residues may be involved in important interactions with other molecules. Furthermore, a person skilled in the art can create test variants containing a single amino acid substitution at each desired amino acid residue. The variants can then be screened using activity assays known to those skilled in the art. Such variants can be used to gather information on suitable variants. For example, if a change in a particular amino acid residue is found to result in impaired, undesirably reduced, or inappropriate activity, then variants with such a change can be avoided. In other words, based on the information obtained from such routine experiments, one skilled in the art can easily determine the amino acid positions at which further substitutions should be avoided, either alone or in combination with other mutations.

As used herein, the terms "polypeptide fragment" and "truncated polypeptide" refer to a polypeptide that has a deletion of the amino terminus and/or carboxy terminus compared to the corresponding full-length protein. In various embodiments, fragments can have a length of, for example, at least 5, at least 10, at least 25, at least 50, at least 100, at least 150, at least 200, at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, at least 600, at least 700, at least 800, at least 900, or at least 1000 amino acids. In various embodiments, the fragments may also have a length of, for example, at most 1000, at most 900, at most 800, at most 700, at most 600, at most 500, at most 450, at most 400, at most 350, at most 300, at most 250, at most 200, at most 150, at most 100, at most 50, at most 25, at most 10, or at most 5 amino acids. The fragment may further comprise at one or both of its ends one or more additional amino acids, for example, an amino acid sequence from another natural protein (e.g., an Fc domain or a leucine zipper domain) or an artificial amino acid sequence (e.g., an artificial linker sequence).

As used herein, the terms "polypeptide variant," "hybrid polypeptide," and "polypeptide mutant" refer to a polypeptide that comprises an amino acid sequence in which one or more amino acid residues are inserted, deleted, and/or substituted in the amino acid sequence compared to another polypeptide sequence. In various embodiments, the number of amino acid residues to be inserted, deleted, or substituted can be, for example, at least 1, at least 2, at least 3, at least 4, at least 5, at least 10, at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 225, at least 250, at least 275, at least 300, at least 350, at least 400, at least 450, or at least 500 amino acids in length. Hybrids according to the description of the present invention include fusion proteins.

A "derivative" of a polypeptide is a polypeptide that has been chemically modified, such as by conjugation with another chemical moiety such as, for example, polyethylene glycol, albumin (e.g., human serum albumin), phosphorylation, and glycosylation.

The term "% sequence identity" is used herein interchangeably with the term "% identity" and refers to the level of amino acid sequence identity between two or more peptide sequences or the level of nucleotide sequence identity between two or more nucleotide sequences when they are aligned using a sequence alignment program. For example, 80% identity as used herein means the same as 80% sequence identity determined using an established algorithm and means that a given sequence is at least 80% identical to another sequence of a different length. In various embodiments, % identity is selected from, for example, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% or more sequence identity with a given sequence. In various embodiments, the % identity ranges from, for example, about 60% to about 70%, about 70% to about 80%, about 80% to about 85%, about 85% to about 90%, about 90% to about 95%, or about 95% to about 99%.

The term "% sequence homology" is used herein interchangeably with the term "% homology" and refers to the level of amino acid sequence homology between two or more peptide sequences or the level of nucleotide sequence identity between two or more nucleotide sequences when they are aligned using a sequence alignment program. For example, 80% homology as used herein means the same as 80% sequence homology determined using an established algorithm, and accordingly, a homologue of a given sequence is characterized by more than 80% sequence homology along the length of the given sequence. In various embodiments, % homology is selected from, for example, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% or more sequence homology with a given sequence. In certain embodiments of the present invention, the % homology is in the range of, for example, from about 60% to about 70%, from about 70% to about 80%, from about 80% to about 85%, from about 85% to about 90%, from about 90% to about 95%, or from about 95% to about 99%.

Examples of computer programs that can be used to determine the identity between two sequences include, but are not limited to, the BLAST suite of programs, such as BLASTN, BLASTX and TBLASTX, BLASTP, and TBLASTN, which are publicly available on the Internet at the NCBI website. See also Altschul et al., J. Mol. Biol. 215: 403–10, 1990 (with special reference to the published default parameters, i.e., w = 4, t = 17) and Altschul et al., Nucleic Acids Res., 25: 3389–3402, 1997. Sequence searches are typically performed using the BLASTP program by evaluating a given amino acid sequence against amino acid sequences in GenBank protein sequences and other publicly available databases. BLASTX is the preferred program for searching nucleic acid sequences translated in all reading frames against amino acid sequences in GenBank protein sequence databases and other publicly available databases. Both BLASTP and BLASTX are run using default parameters: a gap-introduction penalty of 11.0 and a gap-extension penalty of 1.0, and use the BLOSUM-62 matrix.

In addition to calculating the percent sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin & Altschul, Proc. Nat'l. Acad. Sci. USA, 90:5873–5787, 1993). One of the similarity parameters obtained by the BLAST algorithm is the smallest probability sum (P(N)), which indicates the probability of a match between two nucleotide or amino acid sequences occurring by chance. For example, a nucleic acid is considered similar to a reference sequence if the smallest probability sum when comparing the test nucleic acid to the reference nucleic acid is, for example, less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001.

As used herein, the term "modification" refers to any manipulation of the peptide backbone (e.g., amino acid sequence) or post-translational modifications (e.g., glycosylation) of a polypeptide.

As used herein, the term "antigen-binding molecule" refers in its broadest sense to a molecule that specifically binds to an antigenic determinant. Examples of antigen-binding molecules include antibodies, antibody fragments, and proteins that bind to the antigen backbone. "An antigen-binding molecule that binds to the same epitope" as a reference molecule refers to an antigen-binding molecule that blocks the reference molecule from binding to its antigen in a competitive assay by 50% or more, and conversely, the reference molecule blocks the antigen-binding molecule from binding to its antigen in a competitive assay by 50% or more.

As used herein, the term "antigen-binding site" refers to a portion of an antigen-binding molecule that specifically binds to an antigenic determinant. More specifically, the term "antigen-binding site" refers to a portion of an antibody that contains a region that specifically binds to and is complementary to a portion or all of the antigen. When the antigen is large, the antigen-binding molecule may bind only to a specific portion of the antigen, called an epitope. An antigen-binding site may be represented, for example, by one or more variable domains (also called variable regions). Preferably, the antigen-binding site comprises the variable region of the antibody's light chain (VL) and the variable region of the antibody's heavy chain (VH).

As used herein, the term "antigenic determinant" is synonymous with "antigen" and "epitope" and refers to a site (e.g., a continuous sequence of amino acids or a conformational configuration formed by different regions of non-contiguous amino acid sequences) on a polypeptide macromolecule to which an antigen-binding moiety binds, forming an antigen-binding moiety-antigen complex. Useful antigenic determinants can be found, for example, on the surfaces of tumor cells, on the surfaces of cells infected with viruses, on the surfaces of cells affected by other diseases, on the surfaces of immune cells, free in blood plasma and/or in the extracellular matrix (ECM). Unless otherwise specified, proteins useful as antigens herein can be any native form of proteins from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). In various embodiments, the antigen is a human protein.

The term "antibody" is used herein in its broadest sense and encompasses various antibody structures, including, but not limited to, monoclonal antibodies, polyclonal antibodies, monospecific and multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, as long as they exhibit the desired antigen-binding activity.

The term "chimeric" antibody refers to an antibody in which a portion of the heavy and/or light chain is derived from a particular source or species, while the remainder of the heavy and/or light chain is derived from a different source or species.

A "humanized" antibody refers to a chimeric antibody comprising amino acid residues of non-human HVRs (hypervariable regions) and amino acid residues of human FRs (framework regions). In some embodiments, a humanized antibody comprises substantially all of at least one, and typically two, variable domains, in which all or substantially all of the HVRs (e.g., CDRs) correspond to domains of a non-human antibody, and all or substantially all of the FRs correspond to domains of a human antibody. A humanized antibody may optionally comprise at least a fragment of an antibody constant region derived from a human antibody. A "humanized form" of an antibody, such as a non-human antibody, refers to an antibody that has undergone humanization. Other forms of "humanized antibody" encompassed by the present invention are those in which the constant region is further modified or altered relative to that of the parent antibody to achieve properties in accordance with the invention, particularly with respect to Clq binding and/or Fc receptor (FcR) binding.

A "human" antibody is an antibody that has an amino acid sequence that corresponds to that of an antibody produced by a human being or a human cell, or derived from a non-human source, that utilizes human antibody repertoires or other human antibody-encoding sequences. This definition of a human antibody specifically excludes a humanized antibody containing non-human antigen-binding residues.

As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical and/or bind to the same epitope, excluding possible antibody variants, such as those containing naturally occurring mutations or arising during the production of the monoclonal antibody preparation, such variants typically present in minor amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody in a monoclonal antibody preparation is directed against a single determinant on the antigen.

As used herein, the term "monospecific" antibody refers to an antibody that has one or more binding sites, each of which binds to the same epitope on the same antigen. The term "bispecific" means that the antibody is capable of specifically binding to at least two separate antigenic determinants, such as two binding sites, each formed by a pair of the antibody's heavy chain variable domain (VH) and light chain variable domain (VL), which bind to different antigens or to different epitopes on the same antigen. Such a bispecific antibody has a 1+1 format. Other bispecific antibody formats include 2+1 formats (containing two binding sites for a first antigen or epitope and one binding site for a second antigen or epitope) or 2+2 formats (containing two binding sites for a first antigen or epitope and two binding sites for a second antigen or epitope). Typically, a bispecific antibody contains two antigen-binding sites, each specific for a different antigenic determinant.

The term "valent" as used in the present invention refers to the presence of a designated number of binding sites in a molecule that bind to an antigen. The terms "bivalent," "tetravalent," and "hexavalent" themselves refer to the presence of two binding sites, four binding sites, and six binding sites, respectively, in a molecule that bind to an antigen. Bispecific antibodies according to the invention are at least "bivalent" and may be "trivalent" or "multivalent" (e.g., "tetravalent" or "hexavalent"). In various embodiments, antibodies according to the present invention have two or more binding sites and are bispecific. That is, antibodies can be bispecific even in cases where there are more than two binding sites (i.e., the antibody is trivalent or multivalent). In particular, the invention relates to bispecific divalent antibodies having one binding site for each antigen to which they specifically bind.

The terms "full-length antibody," "intact antibody," and "complete antibody" are used interchangeably here to refer to an antibody with a structure substantially similar to that of a native antibody. "Native antibodies" refer to naturally occurring immunoglobulin molecules with a variety of structures. For example, native IgG antibodies are heterotetrameric glycoproteins weighing approximately 150,000 daltons and consisting of two light chains and two heavy chains linked by disulfide bonds. From the N- to the C-terminus, each heavy chain has a variable region (VH), also called the variable heavy domain or heavy chain variable domain, followed by three constant domains (CHI, CH2, and CH3), also called the heavy chain constant region. Similarly, from the N- to the C-terminus, each light chain has a variable region (VL), also called the variable light domain or light chain variable domain, followed by a light chain constant domain (CL), also called the light chain constant region.

The heavy chain of an antibody can belong to one of five types, called alpha (IgA), delta (IgD), epsilon (IgE), gamma (IgG), or mu (IgM), some of which can be further divided into subtypes, such as gamma 1 (IgG1), gamma 2 (IgG2), gamma 3 (IgG3), gamma 4 (IgG4), alpha 1 (IgA1), and alpha 2 (IgA2). The light chain of an antibody can belong to one of two types, called kappa and lambda, based on the amino acid sequence of its constant domain.

"Antibody fragment" refers to a molecule, other than an intact antibody, containing a fragment of an intact antibody that binds an antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies, triabodies, tetrabodies, cross-Fab fragments; linear antibodies; single-chain antibody molecules (e.g., scFv); multispecific antibodies formed from antibody fragments; and single-domain antibodies. For a review of some antibody fragments, see Hudson et al., Nat Med 9, 129-134 (2003). For a review of scFv fragments, see, e.g., Pluckthun, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994); see also WO 93/16185; and U.S. Patent Nos. 5,571,894 and 5,587,458. For a discussion of Fab and F(ab')2 fragments containing salvage receptor binding epitope residues and having an increased half-life in vivo, see U.S. Patent No. 5,869,046. Diabodies are antibody fragments with two antigen-binding sites, which may be bivalent or bispecific, see, e.g., EP 404,097; WO 1993/01161; Hudson et al., Nat Med 9, 129-134 (2003); and Hollinger et al., Proc Natl Acad Sci USA 90, 6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al., Nat Med 9, 129-134 (2003). Single-domain antibodies are antibody fragments comprising all or a fragment of a heavy chain variable domain, or all or a fragment of a light chain variable domain of an antibody. In some embodiments, the single-domain antibody is a human single-domain antibody (Domantis, Inc., Waltham, Mass.; see, e.g., U.S. Patent No. 6,248,516 B1). Additionally, antibody fragments comprise single-chain polypeptides having the characteristics of a VH domain, namely, capable of assembling together with a VL domain, or a VL domain, namely, capable of assembling together with a VH domain to function as an antigen-binding site and thus provide the antigen-binding property of a full-length antibody. Antibody fragments can be produced by a variety of methods, including, but not limited to, proteolytic cleavage of an intact antibody, as well as production using recombinant host cells (e.g., E. coli or phage), as described herein.

Papain digestion of an intact antibody yields two identical antigen-binding fragments, termed "Fab" fragments, each containing the variable domains of the heavy and light chains, as well as the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. Thus, as used herein, the term "Fab fragment" refers to an antibody fragment containing a light chain fragment containing the VL domain and the constant domain of the light chain (CL), and the VH domain and the first constant domain (CH1) of the heavy chain. Fab' fragments differ from Fab fragments by the addition of several residues at the carboxyl terminus of the CH1 domain of the heavy chain, including one or more cysteines of the antibody hinge region. Fab'-SH are Fab' fragments where the cysteine residue(s) of the constant domains bear a free thiol group. Pepsin treatment produces an F(ab') ₂ fragment that has two antigen-binding sites (two Fab fragments) and part of the Fc region.

"A single-chain Fab fragment" or "scFab" is a polypeptide consisting of an antibody heavy chain variable domain (VH), an antibody constant domain 1 (CH1), an antibody light chain variable domain (VL), an antibody light chain constant domain (CL) and a linker, wherein said antibody domains and said linker have one of the following orders in the direction from the N-terminus to the C-terminus: a) VH-CH1-linker-VL-CL, b) VL-CL-linker-VH-CH1, c) VH-CH1-linker-VL-CH1 or d) VL-CH1-linker-VH-CL; and wherein said linker is a polypeptide consisting of at least 30 amino acids, preferably from 32 to 50 amino acids. These single-chain Fab fragments are stabilized by a natural disulfide bond between the CL domain and the CH1 domain. Additionally, these single-chain Fab molecules can be further stabilized by forming interchain disulfide bonds through the incorporation of cysteine residues (e.g., position 44 in the variable region of the heavy chain and position 100 in the variable region of the light chain according to Kabat numbering).

A single-chain variable fragment (scFv) is a fusion protein of the variable regions of the heavy (VH) and light (VL) chains of an antibody, linked by a short linker peptide of ten to approximately 25 amino acids in length. The linker is typically enriched with glycine for flexibility and serine or threonine for solubility and can link the N-terminus of VH to the C-terminus of VL, or vice versa. This protein retains the specificity of the original antibody despite the removal of the constant regions and the introduction of the linker. scFv antibodies, for example, are described in Houston, J. S., Methods in Enzymol. 203 (1991) 46–96. In addition, antibody fragments comprise single-chain polypeptides having the characteristics of a VH domain, namely the ability to assemble together with a VL domain, or a VL domain, namely the ability to assemble together with a VH domain to form a functional antigen-binding molecule and thus providing the antigen-binding property of a full-length antibody.

The term "Fc domain" or "Fc region" is used herein to define the C-terminal region of an antibody heavy chain, which contains at least a fragment of the constant region. The term includes native sequence Fc regions and variant Fc regions. In particular, the Fc region of the human IgG heavy chain extends from Cys226 or Pro230 to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc region may or may not be present. The amino acid sequences of heavy chains are always represented with a C-terminal lysine; however, variants without a C-terminal lysine are included in the invention.

The Fc region of IgG comprises an IgG CH2 domain and an IgG CH3 domain. The "CH2 domain" of the Fc region of human IgG typically extends from the amino acid residue at approximately position 231 to the amino acid residue at approximately position 340. In one embodiment, a carbohydrate chain is attached to the CH2 domain. The CH2 domain here may be a native sequence CH2 domain or a variant CH2 domain. The "CH3 domain" comprises the sequence of residues from the C-terminus to the CH2 domain in the Fc region (i.e., from the amino acid residue at approximately position 341 to the amino acid residue at approximately position 447 in IgG). The CH3 region herein may be a native sequence CH3 domain or a variant CH3 domain (e.g., a CH3 domain with an introduced "bump" in one of its chains and a corresponding introduced "cavity" in its other chain; see U.S. Patent No. 5,821,333, incorporated herein by reference). Such CH3 domain variants may be used to facilitate heterodimerization of two non-identical antibody heavy chains, as described herein. Unless otherwise indicated herein, amino acid residue numbering in the Fc region or constant region is performed according to the EU numbering system, also referred to as the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991.

Knob-in-hole technology is described, for example, in U.S. Patent Nos. 5,731,168; 7,695,936; Ridgway et al., Prot Eng 9, 617-621 (1996); and Carter, J Immunol Meth 248, 7-15 (2001). Typically, the method involves introducing a knob on the surface of a first polypeptide into a corresponding cavity on the surface of a second polypeptide such that the knob can be positioned within the cavity in a manner that promotes heterodimer formation and inhibits homodimer formation. The knobs are constructed by replacing small amino acid side chains on the surface of the first polypeptide with larger ones (e.g., tyrosine or tryptophan). Corresponding cavities of identical or smaller size relative to the knobs are created on the surface of the second polypeptide by replacing large amino acid side chains with smaller ones (e.g., alanine or threonine). The knob and cavity can be obtained by replacing the nucleic acid encoding the polypeptides, for example, by site-directed mutagenesis or by peptide synthesis. In a specific embodiment, the knob modification comprises the amino acid substitution T366W in one of the two Fc domain subunits, and the cavity modification comprises the amino acid substitutions T366S, L368A, and Y407V in the other of the two Fc domain subunits. In another specific embodiment, the Fc domain subunit comprising the knob modification further comprises the amino acid substitution S354C, and the Fc domain subunit comprising the cavity modification further comprises the amino acid substitution Y349C. The introduction of these two cysteine residues results in the formation of a disulfide bridge between the two subunits of the Fc region, thus further stabilizing the dimer (Carter, J Immunol Methods 248, 7-15 (2001)).

The "immunoglobulin Fc equivalent region" is intended to include naturally occurring allelic variants of the immunoglobulin Fc region, as well as variants that carry changes that result in substitutions, insertions, or deletions, but that do not substantially reduce the ability of the immunoglobulin to mediate effector functions (such as antibody-dependent cellular cytotoxicity). For example, one or more amino acids may be deleted from the N-terminus or C-terminus of the immunoglobulin Fc region without significant loss of biological function. Such variants may be selected in accordance with general rules known in the art so as to have minimal effect on activity (see, e.g., Bowie, J. U. et al., Science 247:1306-10 (1990)).

The term "effector functions" refers to the biological activities inherent in the Fc region of an antibody, which may vary depending on the antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), cytokine secretion, immune complex-mediated antigen uptake by antigen-presenting cells, downregulation of cell surface receptors (e.g., the B cell receptor), and B cell activation.

An "activating Fc receptor" is an Fc receptor that, upon interaction with the Fc region of an antibody, triggers signaling events that stimulate the receptor-bearing cell to perform effector functions. Activating Fc receptors include FcγRIIIa (CD16a), FcγRI (CD64), FcγRIIa (CD32), and FcaRI (CD89). A specific activating Fc receptor is human FcγRIIIa (see UniProt accession number P08637, version 141).

A "blocking" antibody or "antagonist" antibody is an antagonist that inhibits or reduces the biological activity of the antigen to which it binds. In some embodiments, blocking antibodies or antagonist antibodies substantially inhibit or completely inhibit the biological activity of the antigen. For example, bispecific antibodies according to the invention block the signaling pathway through TGF-β and activin in such a way as to inhibit the TGF-β/activin-Smad2/3 signaling pathway.

As used herein, "specific binding" means that the binding is selective for the antigen and can be distinguished from unwanted or nonspecific interactions. The ability of an antigen-binding molecule to bind to a specific antigen can be measured using an enzyme-linked immunosorbent assay (ELISA) or other methods familiar to those skilled in the art, such as surface plasmon resonance (SPR) (BIAcore assay) (Liljeblad et al., Glyco J 17, 323–329 (2000)), and conventional binding assays (Heeley, Endocr Res 28, 217–229 (2002)).

As used herein, the terms "affinity" or "binding affinity" refer to the strength of the overall non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (K _D ), which is the ratio of the dissociation and association rate constants (k _off and k _on , respectively). A specific method for measuring affinity is surface plasmon resonance (SPR). As used herein, the term "high affinity" of an antibody refers to an antibody having a Kd of 10 ^-9 M or less, and more specifically 10 ^-10 M or less, for a target antigen. The term "low affinity" of an antibody refers to an antibody having a Kd of 10 ^-8 M or greater. As used herein, the term "decreased binding" refers to a decrease in the affinity of the respective interaction, measured, for example, by SPR. Conversely, "increased binding" refers to an increase in the binding affinity of the respective interaction.

The terms "bispecific antibody comprising a first antigen-binding molecule that specifically binds to activin or an activin-related ligand and a second antigen-binding molecule that specifically binds to TGF-β", "bispecific antibody that specifically binds to activin or an activin-related ligand and TGF-β", "bispecific antigen-binding molecule specific for activin or an activin-related ligand and TGF-β" are used interchangeably herein and refer to a bispecific antibody that is capable of binding to activin or an activin-related ligand and TGF-β with sufficient affinity such that such an antibody is useful as a diagnostic and/or therapeutic agent when targeting activin or an activin-related ligand and TGF-β.

The terms "anti-activin antibody" and "antibody comprising an antigen-binding site that binds to activin" refer to an antibody that is capable of binding to activin, particularly an activin polypeptide expressed on the cell surface, with sufficient affinity such that the antibody is useful as a diagnostic and/or therapeutic agent when targeting activin. In one embodiment, the degree of binding of the anti-activin antibody to an unrelated protein other than activin is less than about 10% of the binding of the antibody to activin, measured, for example, by radioimmunoassay (RIA) or flow cytometry (FACS), or by surface plasmon resonance analysis using a biosensor system such as the Biacore® system. In some embodiments, a molecule that binds to an antigen that binds to human activin has a _KD binding affinity value for binding to human activin, for example, from ^10-8 M to ^10-13 M. In one preferred embodiment, the respective _KD binding affinity values are determined in a surface plasmon resonance assay using the extracellular domain (ECD) of human activin (activin-ECD) for binding affinity to activin. The term "anti-activin antibody" also encompasses bispecific antibodies that are capable of binding to activin and a second antigen.

The terms "anti-TGF-β antibody" and "antibody comprising an antigen-binding site that binds to TGF-β" refer to an antibody that is capable of binding to TGF-β, particularly to a TGF-β polypeptide expressed on a cell surface, with sufficient affinity such that the antibody is useful as a diagnostic and/or therapeutic agent in targeting TGF-β. In one embodiment, the degree of binding of the anti-TGF-β antibody to an unrelated protein other than TGF-β is less than about 10% relative to the binding of the antibody to TGF-β, measured, for example, by radioimmunoassay (RIA) or flow cytometry (FACS), or by surface plasmon resonance analysis using a biosensor system such as the Biacore® system. In some embodiments, an antigen-binding molecule that binds to human TGF-β has a _KD binding affinity for binding to human TGF-β of, for example, ^10-8 M to ^10-13 M. In one preferred embodiment, the corresponding _KD binding affinity is determined in a surface plasmon resonance assay using the extracellular domain (ECD) of human TGF-β (TGF-β-ECD) for TGF-β binding affinity. The term "anti-TGF-β antibody" also encompasses bispecific antibodies capable of binding TGF-β and a second antigen.

As used herein, the term "fusion protein" refers to a fusion polypeptide molecule comprising two or more genes that originally encoded separate proteins, wherein the components of the fusion protein are linked to each other by peptide bonds, either directly or via peptide linkers. As used herein, the term "fusion" refers to components linked by peptide bonds, either directly or via one or more peptide linkers.

A "linker" refers to a molecule that connects two other molecules, either covalently or through ionic, van der Waals, or hydrogen bonds, such as a nucleic acid molecule that hybridizes to one complementary sequence at the 5' end and to another complementary sequence at the 3' end, thus joining two non-complementary sequences. A "cleavable linker" refers to a linker that can be degraded or otherwise separated into two components connected by a cleavable linker. Typically, cleavable linkers are cleaved by enzymes, typically peptidases, proteases, nucleases, lipases, and the like. Cleavable linkers can also be cleaved by environmental factors, such as changes in temperature, pH, salt concentration, and the like.

As used herein, the term "peptide linker" refers to a peptide comprising one or more amino acids, typically approximately 2-20 amino acids. Peptide linkers are known in the art or described herein. Suitable non-immunogenic linker peptides include, for example, ( _G4S ) _n , ( _SG4 ) _n , or _G4 (SG4) _n peptide linkers. "n" is typically a number from 1 to 10, typically from 2 to 4.

"Pharmaceutical composition" refers to a composition suitable for pharmaceutical use in an animal. The pharmaceutical composition comprises a pharmacologically effective amount of an active agent and a pharmaceutically acceptable carrier. "Pharmacologically effective amount" refers to that amount of the agent that is effective in achieving the desired pharmacological result. "Pharmaceutically acceptable carrier" refers to any of the standard pharmaceutical carriers, diluents, buffers, and excipients, such as phosphate-buffered saline, 5% aqueous dextrose solution, and emulsions such as oil-in-water or water-in-oil emulsions, and various types of humectants and/or adjuvants. Suitable pharmaceutical carriers and compositions are described in Remington's Pharmaceutical Sciences, 21st Ed. 2005, Mack Publishing Co, Easton. A "pharmaceutically acceptable salt" is a salt that can be formulated into a compound for pharmaceutical use, including, for example, metal salts (sodium, potassium, magnesium, calcium, etc.) and salts of ammonia or organic amines.

As used herein, "treatment" (and its grammatical variations, such as "treat" or "treatment process") refers to a clinical intervention that attempts to alter the natural course of a disease in the individual being treated and may be performed prophylactically or during the course of clinical pathology. Desired effects of treatment include, but are not limited to, prevention of disease onset or recurrence, alleviation of symptoms, reduction of any direct or indirect pathological consequences of the disease, prevention of metastasis, reduction in the rate of disease progression, amelioration or alleviation of the disease state, and remission or an improved prognosis. As used herein, the term "alleviation" of a disease, disorder, or condition means a reduction in the severity and/or frequency of symptoms of the disease, disorder, or condition. Furthermore, references herein to "treatment" include references to curative, palliative, and prophylactic treatment.

As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of a compound or composition sufficient to treat the specified disorder, condition, or disease, such as ameliorating, alleviating, reducing, and/or delaying one or more of its symptoms. With respect to cancer or other unwanted cellular proliferation, an effective amount includes an amount sufficient to (1) reduce the number of cancer cells; (2) reduce the size of a tumor; (3) inhibit, delay, slow to some extent, and preferably stop the infiltration of cancer cells into peripheral organs; (4) inhibit (i.e., slow to some extent, and preferably stop) tumor metastasis; (5) inhibit tumor growth; (6) prevent or delay the onset and/or recurrence of a tumor; and/or (7) relieve to some extent one or more symptoms associated with cancer. An effective amount may be administered by one or more than one administration.

The phrase "administration" or "procedure for administration" refers to the actions taken by a healthcare professional (e.g., a physician) or a person supervising the care of a patient who supervises and/or authorizes the administration of the agent(s)/compound(s) in question to the patient. Providing administration may include diagnosis and/or determining an appropriate therapeutic regimen and/or prescribing the specific agent(s)/compound(s) to the patient. Such prescribing may include, for example, preparing a prescription form, annotating a medical record, etc. Where administration is described herein, "procedure for administration" is also included.

The terms "patient," "individual," and "subject" may be used interchangeably and refer to a mammal, preferably a human or non-human primate, as well as domesticated mammals (e.g., a dog or cat), laboratory mammals (e.g., a mouse, rat, rabbit, hamster, guinea pig), and farm mammals (e.g., a horse, bovine, swine, sheep). In various embodiments, the patient may be a human (e.g., an adult male, adult female, adolescent male, adolescent female, infant male, infant female) under the care of a physician or other healthcare professional in a hospital, mental health facility, outpatient, or other clinical context. In various embodiments, the patient may be an immunocompromised patient or a patient with a weakened immune system, including, but not limited to, patients suffering from primary immunodeficiency, AIDS; cancer and transplant patients who are taking certain immunosuppressive medications; and patients with inherited diseases that affect the immune system (e.g., congenital agammaglobulinemia, congenital IgA deficiency). In various embodiments, the patient suffers from an immunogenic cancer, including, but not limited to, bladder cancer, lung cancer, melanoma, and other cancers that have been reported to have a high mutation rate (Lawrence et al., Nature, 499(7457):214-218, 2013).

The term "immunotherapy" refers to cancer treatments that include, but are not limited to, treatments using depleting antibodies to specific tumor antigens; treatments using antibody-drug conjugates; treatment with agonist, antagonist, or blocking antibodies against costimulatory or coinhibitory molecules (immune checkpoints) such as CTLA-4, PD-1, OX-40, CD137, GITR (glucocorticoid-induced tumor necrosis factor receptor), LAG3 (leukocyte activation gene 3), TIM-3 (T-cell immunoglobulin and mucin domain-containing molecule 3), SIRP (signal regulatory protein), CD40, CD47, Siglec (sialic acid-binding immunoglobulin-like lectin) 8, Siglec 9, Siglec 15, TIGIT (T-cell immunoreceptor with Ig and ITIM (immunoreceptor tyrosine-linked inhibitory motif) domains), and VISTA (suppressor of T-cell activation containing immunoglobulin V region); Treatment with bispecific T-cell engager antibodies (BiTE®) such as blinatumomab Treatment that includes administration of biological response modifiers such as IL-2 (interleukin), IL-12, IL-15, IL-21, GM-CSF (granulocyte-macrophage column stimulating factor), IFN-α, IFN-β, and IFN-γ Treatment with therapeutic vaccines such as sipuleucel-T Treatment with bacille Calmette-Guérin (BCG) Treatment with dendritic cell vaccines, or tumor antigen-based peptide vaccines Treatment with chimeric antigen receptor (CAR) T cells Treatment with CAR-NK natural killer cells Treatment with tumor infiltrating lymphocytes (TIL) treatment using adoptively transferred anti-tumor T cells (ex vivo dividing and/or TCR transgenic); treatment using TALL-104 cells; and treatment using immunostimulatory agents such as Toll-like receptor (TLR) CpG agonists and imiquimod.

[001] The term "resistant or poorly responsive cancer" refers to tumor cells or cancer that are not responsive to previous anticancer therapy, including, for example, chemotherapy, surgery, radiation therapy, stem cell transplantation, and immunotherapy. Tumor cells may be resistant or poorly responsive at the start of treatment, or they may become resistant or poorly responsive as treatment progresses. Poorly responsive tumor cells include tumor cells that do not respond at the start of treatment or that initially respond for a short period of time but are not responsive to treatment. Poorly responsive tumor cells also include tumor cells that are responsive to treatment using anticancer therapy but are not responsive to subsequent courses of therapy. For the purposes of this invention, difficult-to-treat tumor cells also encompass tumor cells that appear to be suppressed by anticancer therapy but recur within five years, sometimes up to ten years or longer, after treatment is stopped. Anticancer therapy may employ chemotherapy alone, radiation alone, targeted therapy alone, immunotherapy alone, surgery alone, or combinations thereof. For ease of description and without limitation, the terms "difficult-to-treat tumor cells" and "resistant tumor" are used interchangeably.

As used herein, the term "polymer" generally includes, but is not limited to, homopolymers; copolymers, such as block copolymers, graft copolymers, random copolymers, and alternating copolymers; and terpolymers; and blends and modifications thereof. Furthermore, unless specifically limited otherwise, the term "polymer" shall include all possible geometric configurations of the material. Such configurations include, but are not limited to, isotactic, syndiotactic, and random configurations.

"Polynucleotide" refers to a polymer composed of nucleotide monomers. Polynucleotides include naturally occurring nucleic acids such as deoxyribonucleic acid ("DNA") and ribonucleic acid ("RNA"), as well as nucleic acid analogs. Nucleic acid analogs include those that include non-naturally occurring bases, nucleotides that engage in linkages with other nucleotides other than the naturally occurring phosphodiester bond, or that include bases that bind via linkages other than phosphodiester bonds. Thus, nucleotide analogs include, for example and without limitation, phosphorothioates, phosphorodithioates, phosphorotriesters, phosphoramidates, boranophosphates, methylphosphonates, chiral-methylphosphonates, 2-O-methyl ribonucleotides, peptide nucleic acids (PNA), and the like. Such polynucleotides can be synthesized, for example, using an automated DNA synthesizer. The term "nucleic acid" generally refers to large polynucleotides. The term "oligonucleotide" generally refers to short polynucleotides, typically no more than approximately 50 nucleotides long. It is understood that when a nucleotide sequence is represented by a DNA sequence (i.e., A, T, G, C), it also includes an RNA sequence (i.e., A, U, G, C), in which "U" is replaced by "T."

Here, a conventional notation is used to describe polynucleotide sequences: the left end of a single-stranded polynucleotide sequence is the 5' end; the leftward direction of a double-stranded polynucleotide sequence is called the 5' direction. The direction of nucleotide addition from 5' to 3' to nascent RNA transcripts is called the direction of transcription. The strand of DNA that has the same sequence as the mRNA is called the "coding strand"; sequences on the DNA strand that have the same sequence as the mRNA transcribed from it and that are 5' relative to the 5' end of the RNA transcript are called "upstream sequences"; sequences on the DNA strand that have the same sequence as the RNA and that are 3' relative to the 3' end of the coding RNA transcript are called "downstream sequences".

"Complementary" refers to the topological compatibility or mutual fit of the interacting surfaces of two polynucleotides. Thus, two molecules can be described as complementary, and, furthermore, the characteristics of the contact surface are complementary to each other. A first polynucleotide is complementary to a second polynucleotide if the nucleotide sequence of the first polynucleotide is substantially identical to the nucleotide sequence of the polynucleotide-binding partner of the second polynucleotide, or if the first polynucleotide can hybridize to the second polynucleotide under stringent hybridization conditions.

"Hybridization specific for" or "specific hybridization" or "selective hybridization to" refers to the binding, duplex formation, or hybridization of a nucleic acid molecule preferentially to a specific nucleotide sequence under stringent conditions when that sequence is present in a complex mixture (e.g., total cellular) of DNA or RNA. The term "stringent conditions" refers to conditions under which a probe will hybridize preferentially to its target sequence and to a lesser extent to other sequences or to no other sequences. "Stringent hybridization" and "stringent hybridization wash conditions" in the context of nucleic acid hybridization experiments, such as Southern and Northern hybridization, are sequence-dependent and vary under different environmental conditions. Extensive guidance on nucleic acid hybridization can be found in Tijssen, 1993, Laboratory Techniques in Biochemistry and Molecular Biology—Hybridization with Nucleic Acid Probes, part I, chapter 2, "Overview of principles of hybridization and the strategy of nucleic acid probe assays," Elsevier, N.Y.; Sambrook et al., 2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, 3rd sup. rd ed., NY; and Ausubel et al., eds., Current Edition, Current Protocols in Molecular Biology, Greene Publishing Associates and Wiley Interscience, NY.

Typically, highly stringent hybridization and wash conditions are selected to be approximately 5°C lower than the melting point (Tm) temperature for a particular sequence at a given ionic strength and pH. Tm is the temperature (at a given ionic strength and pH) at which 50% of the target sequence hybridizes to a perfectly matched probe. Very stringent conditions are selected to be equal to the Tm for a particular probe. An example of stringent hybridization conditions for hybridization of complementary nucleic acids that have more than approximately 100 complementary residues on the filter in Southern or Northern blotting is 50% formalin with 1 mg heparin at 42°C, with hybridization performed overnight. An example of very stringent wash conditions is 0.15 M NaCl at 72°C for approximately 15 minutes. An example of stringent wash conditions is a wash with 0.2× SSC (sodium citrate and chloride solution) at 65°C for 15 minutes. See Sambrook et al. for a description of SSC buffer. A wash under highly stringent conditions may be preceded by a wash under low stringency conditions to remove background probe signal. A typical moderate-stringency wash for a duplex, for example, greater than approximately 100 nucleotides, is 1× SSC at 45°C for 15 minutes. A typical low-stringency wash for a duplex, for example, greater than approximately 100 nucleotides, is 4-6× SSC at 40°C for 15 minutes. Typically, a signal-to-noise ratio of 2× (or greater) than that detected for an unrelated probe in a particular hybridization assay indicates detection of specific hybridization.

A "primer" refers to a polynucleotide that is capable of specifically hybridizing to a specified polynucleotide template and providing an initiation point for the synthesis of a complementary polynucleotide. Such synthesis occurs when the polynucleotide primer is placed under conditions that induce synthesis, that is, in the presence of nucleotides complementary to the polynucleotide template and a polymerization agent such as DNA polymerase. Primers are typically single-stranded, but may also be double-stranded. Primers are typically deoxyribonucleic acids, but a wide variety of synthetic and natural primers are useful for many applications. A primer is complementary to the template to which it is intended to hybridize to serve as a site for initiation of synthesis, but does not have to mirror the exact sequence of the template. In such a case, specific hybridization of the primer to the template depends on the stringency of the hybridization conditions. Primers can be labeled with, for example, chromogenic, radioactive or fluorescent moieties and used as detectable moieties.

When used in relation to a polynucleotide, a "probe" refers to a polynucleotide capable of specifically hybridizing to a specified sequence of another polynucleotide. A probe specifically hybridizes to a complementary target polynucleotide but need not reflect the exact complementary sequence of the template. In such cases, specific hybridization of the probe to the target depends on the stringency of the hybridization conditions. Probes can be labeled, for example, with chromogenic, radioactive, or fluorescent moieties and used as detectable moieties. In cases where a probe provides an initiation point for the synthesis of a complementary polynucleotide, the probe can also serve as a primer.

A "vector" is a polynucleotide that can be used to introduce another nucleic acid linked to it into a cell. One type of vector is a "plasmid," which refers to a linear or circular double-stranded DNA molecule into which additional nucleic acid segments can be ligated. Another type of vector is a viral vector (e.g., retroviruses, adenoviruses, and replication-defective adeno-associated viruses), in which additional DNA fragments can be introduced into the viral genome. Some vectors are capable of autonomous replication within the host cell into which they are introduced (e.g., bacterial vectors containing a bacterial origin of replication and mammalian episomal vectors). Other vectors (e.g., non-episomal mammalian vectors) integrate into the host cell's genome upon introduction into the host cell and are thus replicated along with the host genome. An "expression vector" is a type of vector that can direct the expression of a selected polynucleotide.

A "regulatory sequence" is a nucleic acid that influences the expression (e.g., the level, timing, or location of expression) of a nucleic acid to which it is operably linked. A regulatory sequence may, for example, exert its effects directly on the regulated nucleic acid or through the action of one or more other molecules (e.g., polypeptides that bind to the regulatory sequence and/or the nucleic acid). Examples of regulatory sequences include promoters, enhancers, and other expression control elements (e.g., polyadenylation signals). Additional examples of regulatory sequences are described, for example, in Goeddel, 1990, Gene Expression Technology Methods in Enzymology 185, Academic Press, San Diego, Calif, and Baron et al., 1995, Nucleic Acids Res. 23:3605–06. A nucleotide sequence is "operably linked" to a regulatory sequence if the regulatory sequence affects the expression (e.g., level, timing, or location of expression) of the nucleotide sequence.

A "host cell" is a cell that can be used to express a polynucleotide as described herein. A host cell can be a prokaryote, such as E. coli, or it can be a eukaryote, such as a single-celled eukaryote (e.g., yeast or another fungus), a plant cell (e.g., a tobacco or tomato cell), an animal cell (e.g., a human cell, a monkey cell, a hamster cell, a rat cell, a mouse cell, or an insect cell), or a hybridoma. Typically, a host cell is a cell cultured in culture that can be transformed or transfected with a nucleic acid encoding a polypeptide, which can then be expressed in the host cell. The phrase "recombinant host cell" can be used to refer to a host cell that has been transformed or transfected with a nucleic acid for expression. A host cell can also be a cell that contains a nucleic acid but does not express it at the desired level, unless a regulatory sequence is introduced into the host cell in such a way that it becomes operably linked to the nucleic acid. It should be understood that the term "host cell" refers not only to a specific cell of the subject but also to the progeny or potential progeny of such a cell. Since certain modifications may occur in subsequent generations due to, for example, mutation or environmental influences, such progeny may not actually be identical to the parent cell, but are still included within the scope of the term as used herein.

The term "isolated molecule" (in cases where the molecule is, for example, a polypeptide or a polynucleotide) is a molecule that, by virtue of its origin or source, (1) is not associated with the naturally associated components that accompany it in its native state, (2) is substantially free of other molecules of the same species, (3) is expressed by a cell of a different species, or (4) is not found in nature. Thus, a molecule that is chemically synthesized or expressed in a cellular system different from the cell from which it naturally originates will be "isolated" from its naturally associated components. A molecule can also be obtained substantially free of naturally associated components by isolation using purification methods well known in the art. The purity or homogeneity of a molecule can be analyzed using a number of methods well known in the art. For example, the purity of a polypeptide sample can be assessed by polyacrylamide gel electrophoresis and staining the gel to visualize the polypeptide using methods well known in the art. For some applications, higher resolution can be achieved using HPLC (high performance liquid chromatography) or other techniques well known in the art for purification.

A protein or polypeptide is "substantially pure," "substantially homogeneous," or "substantially purified" when at least about 60% to 75% of the sample exhibits a single type of polypeptide. The polypeptide or protein may be monomeric or multimeric. A substantially pure polypeptide or protein typically comprises about 50%, 60%, 70%, 80%, or 90% w/w of the protein sample, more preferably about 95%, and will preferably have a purity of greater than 99%. The purity or homogeneity of the protein can be confirmed using a variety of methods well known in the art, such as polyacrylamide gel electrophoresis of the protein sample followed by visualization of a single polypeptide band by staining the gel with a dye well known in the art. For some applications, higher resolution can be achieved using HPLC or other methods well known in the art for purification.

As used herein, the terms "label" or "labeled" refer to the incorporation of another molecule into an antibody. In one embodiment, the label is a detectable marker, such as the incorporation of a radioactively labeled amino acid or the attachment of biotinyl moieties to a polypeptide that can be detected using labeled avidin (e.g., streptavidin containing a fluorescent marker or enzymatic activity that can be detected using optical or calorimetric methods). In another embodiment, the label or marker can be therapeutic, such as a drug conjugate or toxin. Various methods for labeling polypeptides and glycoproteins known in the art can be used. Examples of labels for polypeptides include, but are not limited to, the following: radioisotopes or radionuclides (e.g., ³ H, ¹⁴ C, ¹⁵ N, ³⁵ S, ⁹⁰ Y, ⁹⁹ Tc, ¹¹¹ In, ¹²⁵ I, ¹³¹ I), fluorescent labels (e.g., FITC (fluorescein isothiocyanate), rhodamine, lanthanide phosphors), enzymatic labels (e.g., horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase), chemiluminescent markers, biotinyl groups, predetermined polypeptide epitopes recognized by a secondary reporter (e.g., paired hazel zipper sequences), binding sites for secondary antibodies, metal-binding domains, epitope end groups), magnetic agents such as gadolinium chelates, toxins such as pertussis toxin, taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoside, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol and puromycin and their analogs or homologues. In various embodiments, the labels are attached using spacer arms of varying lengths to reduce potential steric hindrance.

As used herein, the term "heterologous" refers to a composition or state that is not native or found in nature, such as one that can be obtained by replacing an existing natural composition or state with one derived from another source. Similarly, the expression of a protein in an organism different from the organism in which the protein is naturally expressed constitutes a heterologous expression system and a heterologous protein.

It should be understood that aspect and embodiments according to the description of the present invention include “consisting of” and/or “consisting essentially of” aspects and embodiments.

A reference here to "approximately" to a quantity or parameter includes (and describes) variations that address that quantity or parameter as such. For example, a description that refers to "approximately X" includes the description "X."

As used herein and in the claims, the singular also includes the plural unless the context clearly dictates otherwise. It should be understood that aspects and variations according to the invention described herein include "consisting of" and/or "consisting essentially of" aspects and variations.

Activin and activin-related ligands

Activins, including activin A, activin B, and activin AB, and activin-related proteins, including myostatin (GDF-8) and GDF-11, mediate the Smad2/3 signaling pathway by binding to and activating their high-affinity cell surface receptors, ActRIIA and ActRIIB. Activins and related proteins play critical roles in regulating a wide range of biological activities, including mesoderm induction, cellular differentiation, myogenesis, bone remodeling, hematopoiesis, fibrogenesis, and reproductive physiology. Follistatin (FST), a secreted glycoprotein, binds to activins and activin-related ligands to negatively control their signaling activities.

In various embodiments, a bifunctional molecule according to the present invention is capable of binding to activin or an activin-related ligand having an amino acid sequence selected from the group consisting of the amino acid sequences presented in SEQ ID NO: 1-9:

Human ActRIIA-ECD

ETQECLFFNANWEKDRTNQTGVEPCYGDKDKRRHCFATWKNISGSIEIVKQGCWLDDINCYDRTDCVEKKDSPEVYFCCCEGNMCNEKFSYFPEMEVTQPTSNPVTPKPP (SEQ ID NO: 1)

Human ActRIIB-ECD

ETRECIYYNANWELERTNQSGLERCEGEQDKRLHCYASWRNSSGTIELVKKGCWLDDFNCYDRQECVATEENPQVYFCCCEGNFCNERFTHLPEAGGPEVTYEPPPTAPT (SEQ ID NO: 2)

Human follistatin 315

GNCWLRQAKNGRCQVLYKTELSKEECCSTGRLSTSWTEEDVNDNTLFKWMIFNGGAPNCIPCKETCENVDCGPGKKCRMNKKNKPRCVCAPDCSNITWKGPVCGLDGKTYRNECALLKARCKEQPELEVQYQGRCKKTCRDVFCPGSSTCVVDQTNN AYCVTCNRICPEPASSEQYLCGNDGVTYSSACHLRKATCLLGRSIGLAYEGKCIKAKSCEDIQCTGGKKCLWDFKVGRGRCSLCDELCPDSKSDEPVCASDNATYASECAMKEAACSSGVLLEVKHSGSCNSISEDTEEEEEEDEDQDYSFPISSILEW (SEQ ID NO: 3)

Human follistatin AHBS (modified follistatin)

GNCWLRQAKNGRCQVLYKTELSKEECCSTGRLSTSWTEEDVNDNTLFKWMIFNGGAPNCIPCKETCENVDCGPGQSCVVDQTGSPRCVCAPDCSNITWKGPVCGLDGKTYRNECALLKARCKEQPELEVQYQGRCKKTCRDVFCPGSSTCVVDQTNN AYCVTCNRICPEPASSEQYLCGNDGVTYSSACHLRKATCLLGRSIGLAYEGKCIKAKSCEDIQCTGGKKCLWDFKVGRGRCSLCDELCPDSKSDEPVCASDNATYASECAMKEAACSSGVLLEVKHSGSCNSISEDTEEEEEEDEDQDYSFPISSILEW (SEQ ID NO: 4)

Human follistatin 288

GNCWLRQAKNGRCQVLYKTELSKEECCSTGRLSTSWTEEDVNDNTLFKWMIFNGGAPNCIPCKETCENVDCGPGKKCRMNKKNKPRCVCAPDCSNITWKGPVCGLDGKTYRNECALLKARCKEQPELEVQYQGRCKKT CRDVFCPGSSTCVVDQTNNAYCVTCNRICPEPASSEQYLCGNDGVTYSSACHLRKATCLLGRSIGLAYEGKCIKAKSCEDIQCTGGKKCLWDFKVGRGRCSLCDELCPDSKSDEPVCASDNATYASECAMKEAACSSGV LLEVKHSGSCN (SEQ ID NO: 5)

Modified human ActRIIB ECD

ETRECIYYNANWELERTNQSGLERCEGDQDKRLHCYASWRNSSGTIELVKKGCWLDDINCYDRQECVATKENPQVYFCCCEGNFCNERFTHLPEAGGPEVTYEPPPTAPT (SEQ ID NO: 6)

Modified human ActRIIB ECD

ETRECIYYNANWELERTNQSGLERCYGDKDKRRHCYASWRNSSGTIELVKKGCWLDDFNCYDRQECVATEENPQVYFCCCEGNFCNERFTHLPEAGGPEVTYEPPPTAPT (SEQ ID NO: 7)

Modified human ActRIIB ECD

ETRECIYYNANWELERTNQSGLERCEGEQDKRLHCYASWRNSSGTIELVKKGCWDDDFNCYDRQECVATEENPQVYFCCCEGNFCNERFTHLPEAGGPEVTYEPPPT (SEQ ID NO: 8)

Modified human ActRIIA ECD

GAILGRSETQECLFYNANWELERTNQTGVEPCEGEKDKRLHCYATWRNISGSIEIVKKGCWLDDFNCYDRTDCVETEENPQVYFCCCEGNMCNEKFSYFPEMEVTQPTS (SEQ ID NO: 9)

In various embodiments, the multispecific polypeptide molecule is capable of binding to activin or an activin-related ligand having an amino acid sequence selected from the group consisting of the amino acid sequences presented in Table 2:

TGF-β ligands

Transforming growth factor beta (TGF-β), including TGF-β1, TGF-β2, and TGF-β3, mediate the Smad2/3 signaling pathway by binding to and activating the high-affinity cell surface receptors TGFβRII and TGFβRIIB. TGF-β plays a critical role in regulating a wide range of biological activities, including immune function, cell proliferation and differentiation, epithelial-mesenchymal transition, fibrogenesis, hematopoiesis, myogenesis, bone remodeling, cancer progression, and metastasis. Elevated levels of TGF-β and subsequently increased Smad2/3 signaling are implicated in the pathogenesis and development of numerous disease states, including cancer, anemia, bone metastasis, bone loss, fibrosis, pain, muscle wasting, insulin resistance, chronic kidney disease, liver disease, and cardiovascular disease.

In various embodiments, the multispecific polypeptide molecule according to the present invention is capable of binding to a TGF-β ligand having an amino acid sequence selected from the group consisting of the amino acid sequences presented in SEQ ID NO: 18-21:

Human TGF-β receptor II isoform 1

MGRGLLRGLWPLHIVLWTRIASTIPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSD ECNDNIIFSEEYNTSNPDLLLVIFQVTGISLLPPLGVAISVIIIFYCYRVNRQQKLSSTWETGKTRKLMEFSEHCAIILEDDRSDISSTCANNINHNTELLPIELDTLVGKGRFAEVYKAKLKQNTSEQFETVAVKIFPYEE YASWKTEKDIFSDINLKHENILQFLTAEERKTELGKQYWLITAFHAKGNLQEYLTRHVISWEDLRKLGSSLARGIAHLHSDHTPCGRPKMPIVHRDLKSSNILVKNDLTCCLCDFGLSLRLDPTLSVDDLANSGQVGTARYM APEVLESRMNLENVESFKQTDVYSMALVLWEMTSRCNAVGEVKDYEPPFGSKVREHPCVESMKDNVLRDRGRPEIPSFWLNHQGIQMVCETLTECWDHDPEARLTAQCVAERFSELEHLDRLSGRSCSEEKIPEDGSLNTTK (SEQ ID NO: 18)

Human TGF-β receptor isoform 1 II-ECD (TGF-β RIIB-ECD)

TIPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD (SEQ ID NO: 19)

Human TGF-β receptor II isoform 2

MGRGLLRGLWPLHIVLWTRIASTIPPHVQKSDVEMEAQKDEIICPSCNRTAHPLRHINNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAAS PKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPDLLLVIFQVTGISLLPPLGVAISVIIIFYCYRVNRQQKLSSTWETGKTRKLMEFSEHCAIILEDDRSDISSTCANNINHNTELLPIELDTLVGKGRFAEVYKAK LKQNTSEQFETVAVKIFPYEEYASWKTEKDIFSDINLKHENILQFLTAEERKTELGKQYWLITAFHAKGNLQEYLTRHVISWEDLRKLGSSLARGIAHLHSDHTPCGRPKMPIVHRDLKSSNILVKNDLTCCLCDFGLSLRLD PTLSVDDLANSGQVGTARYMAPEVLESRMNLENVESFKQTDVYSMALVLWEMTSRCNAVGEVKDYEPPFGSKVREHPCVESMKDNVLRDRGRPEIPSFWLNHQGIQMVCETLTECWDHDPEARLTAQCVAERFSELEHLDRLSG RSCSEEKIPEDGSLNTTK (SEQ ID NO: 20)

Human TGF-β receptor isoform 2 II-ECD (TGF-β RIIA-ECD)

TIPPHVQKSDVEMEAQKDEIICPSCNRTAHPLRHINNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPD (SEQ ID NO: 21)

In various embodiments, the multispecific polypeptide molecule is capable of binding to a TGF-β ligand having an amino acid sequence selected from the group consisting of the amino acid sequences presented in Table 3:

Antibodies against activin and/or TGF-β and antibody fragments

Methods for producing novel antibodies that bind to activin or an activin-related ligand and/or TGF-β ligands and/or receptors are known to those skilled in the art. For example, a method for producing a monoclonal antibody that specifically binds to activin or an activin-related ligand and/or a TGF-β ligand may include administering to a mouse an immunogenic composition comprising activin or an activin-related ligand and/or a TGF-β ligand in an amount effective to stimulate a detectable immune response, obtaining antibody-producing cells (e.g., cells from the spleen) from the mouse and fusing the antibody-producing cells with myeloma cells to produce antibody-producing hybridomas, and testing the antibody-producing hybridomas to identify a hybridoma that produces a monoclonal antibody that specifically binds to activin or an activin-related ligand and/or a TGF-β ligand. Once obtained, the hybridoma can be expanded into cell culture, possibly under culture conditions where the hybridoma cells produce a monoclonal antibody that specifically binds to activin or an activin-related ligand, and/or a TGF-β ligand. The monoclonal antibody can be purified from the culture. A variety of antigen/antibody interaction testing methods have since become available to identify specific desired antibodies.

Other suitable methods for producing or isolating antibodies with the desired specificity may be used, including, for example, methods that select a recombinant antibody from a library or that rely on immunization of transgenic animals (e.g., mice) capable of producing a complete repertoire of human antibodies. See, for example, Jakobovits et al., Proc. Natl. Acad. Sci. (U.S.A.), 90: 2551-2555, 1993; Jakobovits et al., Nature, 362: 255-258, 1993; Lonberg et al., U.S. Patent No. 5,545,806; and Surani et al., U.S. Patent No. 5,545,807.

Antibodies can be engineered in a variety of ways. They can be produced as single-chain antibodies (including small modular immunopharmaceuticals™ or SMIPs™), Fab and F(ab')2 fragments, etc. Antibodies can be humanized, chimerized, deimmunized, or fully human. Numerous publications describe many types of antibodies and methods for engineering such antibodies. For example, see U.S. Patent Nos. 6,355,245; 6,180,370; 5,693,762; 6,407,213; 6,548,640; 5,565,332; 5,225,539; 6,103,889; and 5,260,203.

Chimeric antibodies can be produced using recombinant DNA technologies known in the art. For example, the gene encoding the Fc constant region of a mouse (or other species) monoclonal antibody molecule is digested with restriction enzymes to remove the mouse Fc coding region and replaced with an equivalent fragment of the gene encoding the human Fc constant region (see Robinson et al., International Patent Application PCT/US86/02269; Akira, et al., European Patent Application 184187; Taniguchi, M., European Patent Application 171496; Morrison et al., European Patent Application 173494; Neuberger et al., International Patent Application WO 86/01533; Cabilly et al., U.S. Patent No. 4,816,567; Cabilly et al., European Patent Application 125,023; Better et al., Science, 240:1041-1043, 1988; Liu et al., Proc. Natl. Acad. Sci. (U.S.A.), 84:3439-3443, 1987; Liu et al., J. Immunol., 139:3521-3526, 1987; Sun et al., Proc. Natl. Acad. Sci. (U.S.A.), 84:214-218, 1987; Nishimura et al., Cane. Res., 47:999-1005, 1987; Wood et al., Nature, 314:446-449, 1985; and Shaw et al., J. Natl Cancer Inst, 80:1553-1559, 1988).

Methods for humanizing antibodies are well known in the art. In some embodiments, a humanized antibody has one or more amino acid residues introduced into it from a source that is non-human, in addition to the non-human CDRs. Humanization can essentially be accomplished according to the method developed by Winter et al. (Jones et al., Nature, 321:522-525, 1986; Riechmann et al., Nature, 332:323-327, 1988; Verhoeyen et al., Science, 239:1534-1536, 1988), by replacing the hypervariable region sequences with the corresponding sequences of a human antibody. Accordingly, such "humanized" antibodies are chimeric antibodies (U.S. Patent No. 4,816,567), in which substantially less than the entire intact human variable region is replaced with the corresponding sequence from a non-human species. In practice, humanized antibodies typically represent human antibodies in which some hypervariable region residues and possibly some framework region residues are replaced with residues from analogous regions of rodent antibodies.

U.S. Patent No. 5,693,761 to Queen et al. discloses subtleties consistent with Winter et al. regarding humanized antibodies and is based on the premise that loss of avidity is attributed to problems in structural motifs in the humanized framework region, which, due to steric or other chemical incompatibility, interfere with the folding of the CDRs into the binding conformation found in the mouse antibody. To address this issue, Queen advocates the use of human framework sequences that are highly homologous in linear peptide sequence to the framework sequences of the mouse antibody to be humanized. Accordingly, Queen's methods focus on comparing framework sequences across species. Typically, all available human variable region sequences are compared to a specific mouse sequence, and the percent identity between corresponding framework residues is calculated. The human variable region with the highest percent identity is selected to generate the framework sequences for humanization. Queen also emphasizes the importance of retaining certain amino acid residues from the mouse framework in the humanized framework, which are critical for maintaining the CDR in a binding-capable conformation. Potential criticality is assessed using molecular models. Typically, candidate residues for retention are those located in a linear sequence near the CDR region or physically within 6 Å (0.6 nm) of any CDR residue.

Another method for antibody humanization, called "framework shuffling," relies on the creation of a combinatorial library with non-human CDR variable regions fused in frame to a collection of individual human terminal framework regions (Dall'Acqua et al., Methods, 36:43, 2005). The libraries are then screened to identify clones encoding humanized antibodies that retain good binding.

Methods for producing fully human antibodies are described in the art. As an example, a method for producing an antibody against activin or an antigen-binding fragment thereof includes the steps of synthesizing a library of human antibodies on phage, screening the library with an activin polypeptide or an antibody-binding fragment thereof, isolating phage that binds the activin polypeptide, and producing the antibody from the phage. As another example, one method for preparing a library of antibodies for use in phage display methods includes the steps of immunizing a non-human animal containing human immunoglobulin loci with an activin polypeptide or an antigen fragment thereof to elicit an immune response, isolating antibody-producing cells from the immunized animal; isolating RNA encoding the heavy and light chains of the antibodies according to the invention from the isolated cells, reverse transcribing the RNA to produce cDNA, amplifying the cDNA using primers, and inserting the cDNA into a phage display vector such that the antibodies are expressed on the phage. In this way, recombinant antibodies against activin can be obtained according to the invention.

Recombinant human antibodies against activin and/or TGF-β according to the invention can also be isolated by screening a combinatorial library of recombinant antibodies. Preferably, the library is a phage display scFv library generated using human V _L and V _H cDNAs derived from mRNA isolated from B cells. Methods for generating and screening such libraries are known in the art. Kits for generating phage display libraries are commercially available (e.g., the Pharmacia Recombinant Phage Antibody System, catalog no. 27-9400-01; and the Stratagene SurfZAP™ phage display kit, catalog no. 240612). There are also other methods and reagents that can be used to generate and screen antibody display libraries (see, e.g., U.S. Patent No. 5,223,409; PCT Publication Nos. WO 92/18619, WO 91/17271, WO 92/20791, WO 92/15679, WO 93/01288, WO 92/01047, WO 92/09690; Fuchs et al., Bio/Technology, 9:1370–1372 (1991); Hay et al., Hum. Antibod. Hybridomas, 3:81–85, 1992; Huse et al., Science, 246:1275–1281, 1989; McCafferty et al., Nature, 348:552-554, 1990; Griffiths et al., EMBO J., 12:725-734, 1993; Hawkins et al., j. Mol. Biol., 226:889-896, 1992; Clackson et al., Nature, 352:624-628, 1991; Gram et al., Proc. Natl. Acad. Sci. (USA), 89:3576-3580, 1992; Garrad et al., Bio/Technology, 9:1373-1377, 1991; Hoogenboom et al., Nuc. Acid Res., 19:4133-4137, 1991; and Barbas et al., Proc. Natl. Acad. Sci. (USA), 88:7978-7982, 1991), incorporated herein by reference.

Human antibodies are also produced by immunizing a transgenic non-human animal that contains some or all of the human immunoglobulin heavy chain and light chain loci with human IgE antigen, such as the XenoMouse™ animal (Abgenix, Inc./Amgen, Inc.--Fremont, Calif.). XenoMouse™ mice are engineered mouse strains that contain large fragments of the human immunoglobulin heavy chain and light chain loci and are defective in mouse antibody production. See, for example, Green et al., Nature Genetics, 7:13-21, 1994 and U.S. Patents 5,916,771, 5,939,598, 5,985,615, 5,998,209, 6,075,181, 6,091,001, 6,114,598, 6,130,364, 6,162,963, and 6,150,584. XenoMouse™ mice produce an adult human repertoire of fully human antibodies and produce antigen-specific human antibodies. In some embodiments, XenoMouse™ mice contain approximately 80% of the human antibody V gene repertoire by introducing megabase-sized fragments of the terminal configuration of the human heavy chain loci and the kappa light chain loci into a yeast artificial chromosome (YAC). In other embodiments, the XenoMouse™ mice further comprise approximately the entire human lambda light chain locus. See Mendez et al., Nature Genetics, 15:146-156, 1997; Green and Jakobovits, J. Exp. Med., 188:483-495, 1998; and WO 98/24893. In one aspect, the present invention provides a method for producing anti-activin and/or TGF-β antibodies from non-human and non-mouse animals by immunizing non-human transgenic animals that comprise human immunoglobulin loci with an activin and/or TGF-β polypeptide. Such animals can be produced using the methods described in the aforementioned documents.

Antibodies against activin

In various embodiments of the present invention, the anti-activin antibody is an anti-activin A antibody that is a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 10:

QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGLSWVRQAPGQGLEWMGWIIPYNGNTNSAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYFCARDRDYGVNYDAFDIWG QGTMVTVSSASTKGPSVFPPLAPSSКSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNVYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 10)

a human antibody or antigen-binding fragment comprising the light chain amino acid sequence set forth in SEQ ID NO: 11:

SYEVTQAPSVSVSPGQTASITCSGDKLGDKYACWYQQKPGQSPVLVIYQDSKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQAWDSSTAVFGGGTKLTVL RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 11)

or a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence shown in SEQ ID NO: 10 and the light chain amino acid sequence shown in SEQ ID NO: 11;

a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region as set forth in SEQ ID NO: 12:

QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGLSWVRQAPGQGLEWMGWIIPYNGNTNSAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYFCARDRDYGVNYDAFDIWGQGTMVTVSS (SEQ ID NO: 12)

a human antibody or antigen-binding fragment comprising the amino acid sequence of a light chain variable region as set forth in SEQ ID NO: 13:

SYEVTQAPSVSVSPGQTASITCSGDKLGDKYACWYQQKPGQSPVLVIYQDSKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQAWDSSTAVFGGGTKLTVL (SEQ ID NO: 13)

or a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 12 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 13.

In various embodiments of the invention, antibodies are provided that comprise a heavy chain, a light chain, or both a heavy chain and a light chain; a heavy chain variable region, a light chain variable region, or both a heavy chain variable region and a light chain variable region; wherein the heavy chain, light chain, heavy chain variable region, or light chain variable region comprises a sequence that has at least 75%, at least about 80%, at least about 85%, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least about 99% identity to the amino acid sequences set forth in SEQ ID NO: 10, 11, 12, or 13; wherein the antibody specifically binds to human activin A.

In various embodiments of the present invention, the anti-activin antibody is an anti-activin A antibody that is a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 14:

QVQLQESGPGLVKPSETLSLTCTVSGGSFSSHFWSWIRQPPGKGLEWIGYILYTGGTSFNPSLKSRVSMSVGTSKNQFSLKLSSVTAADTAVYYCARARSGITFTGIIVPGSF DIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVES KYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIE KTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 14)

a human antibody or antigen-binding fragment comprising the light chain amino acid sequence shown in SEQ ID NO: 15:

EI VLTQS PGTLS LS PGERATLSCR ASQS VS S S YLAW YQQKPGQ APRLLIYGAS S RATGIPDRFSGSGGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KD S T YS LS S TLTLS K AD YEKHK V Y ACE VTHQGLS S P VTK S FNRGEC (SEQ ID NO: 15)

or a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence shown in SEQ ID NO: 14 and the light chain amino acid sequence shown in SEQ ID NO: 15;

a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region as set forth in SEQ ID NO: 16:

QVQLQESGPGLVKPSETLSLTCTVSGGSFSSHFWSWIRQPPGKGLEWIGYILYT GGTS FNP S LKS RVS MS VGTS KNQFS LKLS S VT AADTA VYYC ARARS GITFTGII VPGSF DIWGQGTMVTVSS (SEQ ID NO: 16)

a human antibody or antigen-binding fragment comprising the amino acid sequence of a light chain variable region as set forth in SEQ ID NO: 17:

EI VLTQS PGTLS LS PGERATLSCR ASQS VS S S YLAW YQQKPGQ APRLLIYGAS S RATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIK (SEQ ID NO: 17)

or a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 16 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 17.

In various embodiments of the invention, antibodies are provided that comprise a heavy chain, a light chain, or both a heavy chain and a light chain; a heavy chain variable region, a light chain variable region, or both a heavy chain variable region and a light chain variable region; wherein the heavy chain, light chain, heavy chain variable region, or light chain variable region comprises a sequence that has at least 75%, at least about 80%, at least about 85%, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least about 99% identity to the amino acid sequences set forth in SEQ ID NO: 14, 15, 16, or 17; wherein the antibody specifically binds to human activin A.

Antibodies against TGF-β

In various embodiments of the present invention, the anti-TGF-β antibody is an anti-TGF-β antibody that is a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 22:

QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQGLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRSEDTAVYYCASTLGLVLDAMDYWG QGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYG PPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 22)

a human antibody or antigen-binding fragment comprising the light chain amino acid sequence shown in SEQ ID NO: 23:

ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQAPRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYADS PITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH KVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 23)

or a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence shown in SEQ ID NO: 22 and the light chain amino acid sequence shown in SEQ ID NO: 23;

a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region as set forth in SEQ ID NO: 24:

QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQGLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRSEDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS (SEQ ID NO: 24)

a human antibody or antigen-binding fragment comprising the amino acid sequence of a light chain variable region as set forth in SEQ ID NO: 25:

ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQAPRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYADSPITFGQGTRLEIK (SEQ ID NO: 25)

or a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 24 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 25.

In various embodiments of the invention, antibodies are provided that comprise a heavy chain, a light chain, or both a heavy chain and a light chain; a heavy chain variable region, a light chain variable region, or both a heavy chain variable region and a light chain variable region; wherein the heavy chain, light chain, heavy chain variable region, or light chain variable region comprises a sequence that has at least 75%, at least about 80%, at least about 85%, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least about 99% identity to the amino acid sequences set forth in SEQ ID NO: 22, 23, 24, or 25; wherein the antibody specifically binds to human TGF-β.

PD-1 and PD-L1 and CTLA-4 ligands

Multiple immune checkpoint protein antigens have been reported to be overexpressed on various immune cells, including, for example, SIRP (expressed on macrophages, monocytes, dendritic cells), CD47 (highly expressed on tumor cells and other cell types), VISTA (expressed on monocytes, dendritic cells, B cells, T cells), CD152 (expressed by activated CD8+ T cells, CD4+ T cells, and regulatory T cells), CD279 (expressed on tumor-infiltrating lymphocytes, expressed by activated T cells (both CD4 and CD8), regulatory T cells, activated B cells, activated NK cells, anergic T cells, monocytes, dendritic cells), CD274 (expressed on T cells, B cells, dendritic cells, macrophages, vascular endothelial cells, and islet cells) and CD223 (expressed on activated T cells, regulatory T cells, anergic T cells, NK cells, NKT cells, and plasmacytoid dendritic cells) (see, e.g., Pardoll, D., Nature Reviews Cancer, 12:252-264, 2012). Antibodies that bind to an antigen that has been identified as an immune checkpoint protein are known to those skilled in the art. For example, various antibodies against CD276 have been described in the art (see, e.g., U.S. Patent No. 20120294796 (Johnson et al) and the references cited therein); Anti-CD272 antibodies have been described in the art (see, e.g., U.S. Patent No. 20140017255 (Mataraza et al) and references cited therein); various anti-CD152/CTLA-4 antibodies have been described in the art (see, e.g., U.S. Patent No. 20130136749 (Korman et al) and references cited therein); various anti-LAG-3/CD223 antibodies have been described in the art (see, e.g., U.S. Patent No. 20110150892 (Thudium et al) and references cited therein); various anti-CD279 (PD-1) antibodies have been described in the art (see, e.g., U.S. Patent No. 7488802 (Collins et al) and references cited therein); Various antibodies against CD274 (PD-L1) have been described in the art (see, e.g., U.S. Patent No. 20130122014 (Korman et al.) and references cited therein); various antibodies against TIM-3 have been described in the art (see, e.g., U.S. Patent No. 20140044728 (Takayanagi et al.) and references cited therein); and various antibodies against B7-H4 have been described in the art (see, e.g., U.S. Patent No. 20110085970 (Terrett et al.) and references cited therein); and various antibodies against TIGIT have been described in the art (see, e.g., U.S. Patent No. 20180169239A1 (Grogan) and references cited therein). Each of these references is incorporated herein in its entirety for the particular antibodies and sequences cited therein. In various embodiments, the bifunctional multispecific antagonist molecule is capable of binding to an immune checkpoint protein ligand selected from the group consisting of, but not limited to, CD279 (PD-1), CD274 (PDL-1), CD276, CD272, CD 152, CD223 (LAG-3), CD40, SIRPα, CD47, OX-40, GITR, ICOS, CD27, 4-1BB, TIM-3, B7-H3, B7-H4, TIGIT, and VISTA.

In various embodiments, the bifunctional multispecific antagonist molecule is capable of binding to a PD-1 ligand, a PD-L1 ligand, or a CTLA-4 ligand having an amino acid sequence selected from the group consisting of the amino acid sequences presented in Table 4:

Anti-PD-1, anti-PD-L1, and anti-CTLA-4 antibodies

In various embodiments of the present invention, the anti-PD-1 antibody is an anti-PD-1 antibody that is a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 26:

QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEK TIS K AKGQPREPQ V YTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 26)

a human antibody or antigen-binding fragment comprising the light chain amino acid sequence shown in SEQ ID NO: 27:

ЕIVLTQ S Р ATLS LS PGER ATLSCR ASKG VS TS GY S YLHW YQQKP GQ APRLLIYL AS YLES GVP ARFS GS GS GTDFTLTIS S LEP EDF А V Y YCQHS RDLP LTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 27)

or a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence shown in SEQ ID NO: 26 and the light chain amino acid sequence shown in SEQ ID NO: 27;

a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region as set forth in SEQ ID NO: 28:

QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSS (SEQ ID NO: 28)

a human antibody or antigen-binding fragment comprising the amino acid sequence of a light chain variable region as set forth in SEQ ID NO: 29:

ЕI VLTQ S Р ATLS LS PGER ATLSCR ASKG VS TS GY S YLHW YQQKP GQ APRLLI YL AS YLES GVP ARFS GS GS GTDFTLTIS S LEP EDF A VY YCQHS RDLP LTFGGGTK VEIK (SEQ ID NO: 29)

or a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region as set forth in SEQ ID NO: 28 and the amino acid sequence of the light chain variable region as set forth in SEQ ID NO: 29.

In various embodiments of the invention, antibodies are provided that comprise a heavy chain, a light chain, or both a heavy chain and a light chain; a heavy chain variable region, a light chain variable region, or both a heavy chain variable region and a light chain variable region; wherein the heavy chain, light chain, heavy chain variable region, or light chain variable region comprises a sequence that has at least 75%, at least about 80%, at least about 85%, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least about 99% identity to the amino acid sequences set forth in SEQ ID NO: 26, 27, 28, or 29; wherein the antibody specifically binds to human PD-1.

In various embodiments of the present invention, the anti-PD-1 antibody is an anti-PD-1 antibody that is a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 30:

QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVT VSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTIS KAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 30)

a human antibody or antigen-binding fragment comprising the light chain amino acid sequence shown in SEQ ID NO: 31:

EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNR ATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD S TYS LS S TLTLS K AD YEKHK V Y ACE VTHQGLS SP VTK SFNRGEC (SEQ ID NO: 31)

or a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence shown in SEQ ID NO: 30 and the light chain amino acid sequence shown in SEQ ID NO: 31;

a human antibody or antigen-binding fragment comprising the amino acid sequence of a light chain variable region as set forth in SEQ ID NO: 32:

QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSS (SEQ ID NO: 32

a human antibody or antigen-binding fragment comprising the amino acid sequence of a light chain variable region as set forth in SEQ ID NO: 33

EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNR ATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIK (SEQ ID NO: 33)

or a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 32 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 33.

In various embodiments of the invention, antibodies are provided that comprise a heavy chain, a light chain, or both a heavy chain and a light chain; a heavy chain variable region, a light chain variable region, or both a heavy chain variable region and a light chain variable region; wherein the heavy chain, light chain, heavy chain variable region, or light chain variable region comprises a sequence that has at least 75%, at least about 80%, at least about 85%, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least about 99% identity to the amino acid sequences set forth in SEQ ID NO: 30, 31, 32, or 33; wherein the antibody specifically binds to human PD-1.

In various embodiments of the present invention, the anti-PD-1 antibody is an anti-PD-1 antibody that is a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 38:

EVQLLESGGVLVQPGGSLRLSCAASGTFFSNFGMTWVRQAPGKGLEWVSGISGGGRDTYFADSVKGRFTISRDNSKNTLYLQMNSLKGEDTAVYYCVKWGNIYFDYW GQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRV ESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALH NHYTQKSLSLSLGK (SEQ ID NO: 38)

a human antibody or antigen-binding fragment comprising the light chain amino acid sequence shown in SEQ ID NO: 39:

DIQMTQSPSSLSAS VGDSITITCR AS LSINTFLN W YQQKPGK APNLLIY A AS S LHGGVPSRFSGSGSGTDFTLTIRTLQPEDFATYYCQQSSNTPFTFGPGTVVDFRRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 39)

or a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence shown in SEQ ID NO: 38 and the light chain amino acid sequence shown in SEQ ID NO: 39;

a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region as set forth in SEQ ID NO: 40:

EVQLLESGGVLVQPGGSLRLSCAASGTFFSNFGMTWVRQAPGKGLEWVSGIS GGGRDTYFADSVKGRFTISRDNSKNTLYLQMNSLKGEDTAVYYCVKWGNIYFDYW GQGTLVTVSS (SEQ ID NO: 40)

a human antibody or antigen-binding fragment comprising the amino acid sequence of a light chain variable region as set forth in SEQ ID NO: 41:

DIQMTQSPSSLSAS VGDS ITITCR AS LS INTFLN W YQQKPGK APNLLI Y A AS S LH GGVPSRFSGSGSGTDFTLTIRTLQPEDFATYYCQQSSNTPFTFGPGTVVDFR (SEQ ID NO: 41)

or a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 40 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 41.

In various embodiments of the invention, antibodies are provided that comprise a heavy chain, a light chain, or both a heavy chain and a light chain; a heavy chain variable region, a light chain variable region, or both a heavy chain variable region and a light chain variable region; wherein the heavy chain, light chain, heavy chain variable region, or light chain variable region comprises a sequence that has at least 75%, at least about 80%, at least about 85%, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least about 99% identity to the amino acid sequences set forth in SEQ ID NO: 38, 39, 40, or 41; wherein the antibody specifically binds to human PD-1.

In various embodiments of the present invention, the anti-PD-1 antibody is an anti-PD-1 antibody that is a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 42:

EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYIMMWVRQAPGKGLEWVSSIYPS GGITF Y ADT VKGRFTIS RDN S KNTLYLQMNS LR AEDT A V Y YC ARIKLGT VTT VD YW GQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 42)

a human antibody or antigen-binding fragment comprising the light chain amino acid sequence shown in SEQ ID NO: 43:

QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDV SNRP SGVSNRF SGS KSGNT AS LTIS GLQ AEDE AD Y YC S S YTS S S TR VFGTGTK VTVLG QPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKP SKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 43)

or a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence shown in SEQ ID NO: 42 and the light chain amino acid sequence shown in SEQ ID NO: 43;

a human antibody or antigen-binding fragment comprising the amino acid sequence of a light chain variable region as set forth in SEQ ID NO: 44

EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYIMMWVRQAPGKGLEWVSSIYP S GGITF Y ADT VKGRFTIS RDN S KNTLYLQMNS LR AEDT A V Y YC ARIKLGT VTT VD YW GQGTLVTVSS (SEQ ID NO: 44)

a human antibody or antigen-binding fragment comprising the amino acid sequence of a light chain variable region as set forth in SEQ ID NO: 45:

QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDV SNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYTSSSTRVFGTGTKVTVL (SEQ ID NO: 45)

or a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region as set forth in SEQ ID NO: 44 and the amino acid sequence of the light chain variable region as set forth in SEQ ID NO: 45.

In various embodiments of the invention, antibodies are provided that comprise a heavy chain, a light chain, or both a heavy chain and a light chain; a heavy chain variable region, a light chain variable region, or both a heavy chain variable region and a light chain variable region; wherein the heavy chain, light chain, heavy chain variable region, or light chain variable region comprises a sequence that has at least 75%, at least about 80%, at least about 85%, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least about 99% identity to the amino acid sequences set forth in SEQ ID NO: 42, 43, 44, or 45; wherein the antibody specifically binds to human PD-1.

In various embodiments of the present invention, the anti-CTLA-4 antibody is an anti-CTLA-4 antibody that is a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 34:

QVQLVESGGGVVQPGRSLRLSCAASGFFSSYTMHWVRQAPGKGLEWVTFIS YDGNNK Y Y ADS VKGRFTIS RDN S KNTLY LQMN S LR AEDT AIY YC ARTGWLGPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTP A V LQS S GLY S LS S V VT VP S S S LGTQT YICN VNHKP SNTK VDKR VEPK S CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVFTNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPI EKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 34)

a human antibody or antigen-binding fragment comprising the light chain amino acid sequence shown in SEQ ID NO: 35:

EIVLTQSPGTLSLSPGERATLSCRASQSVGSSYLAWYQQKPGQAPRLLIYGAFS RATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KD S T YS LS S TLTLS K AD YEKHK V Y ACE VTHQGLS S P VTK S FNRGEC (SEQ ID NO: 35)

or a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence shown in SEQ ID NO: 34 and the light chain amino acid sequence shown in SEQ ID NO: 35;

a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region as set forth in SEQ ID NO: 36:

QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYTMHWVRQAPGKGLEWVTFIS YDGNNK Y Y ADS VKGRFTIS RDN S KNTLYLQMNS LR AED T Al Y YC ARTGW LGPFD Y WGQGTLVTVSS (SEQ ID NO: 36)

a human antibody or antigen-binding fragment comprising the amino acid sequence of a light chain variable region as set forth in SEQ ID NO: 37:

EIVLTQSPGTLSLSPGERATLSCRASQSVGSSYLAWYQQKPGQAPRLLIYGAFS RATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIK (SEQ ID NO: 37)

or a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 36 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 37.

In various embodiments of the invention, antibodies are provided that comprise a heavy chain, a light chain, or both a heavy chain and a light chain; a heavy chain variable region, a light chain variable region, or both a heavy chain variable region and a light chain variable region; wherein the heavy chain, light chain, heavy chain variable region, or light chain variable region comprises a sequence that has at least 75%, at least about 80%, at least about 85%, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least about 99% identity to the amino acid sequences set forth in SEQ ID NO: 34, 35, 36, or 37; wherein the antibody specifically binds to human CTLA-4.

In various embodiments of the present invention, the anti-PD-L1 antibody is an anti-PD-L1 antibody that is a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 46:

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWIS PYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDY WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGAL TS G VHTFP A V LQS S GLY S LS S V VT VP S S S LGTQT YICN VNHKP SNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 46)

a human antibody or antigen-binding fragment comprising the light chain amino acid sequence shown in SEQ ID NO: 47:

DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASF LYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KD S T YS LS S TLTLS K AD YEKHK V Y ACE VTHQGLS S P VTK S FNRGEC (SEQ ID NO: 47)

or a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence shown in SEQ ID NO: 46 and the light chain amino acid sequence shown in SEQ ID NO: 47;

a human antibody or antigen-binding fragment comprising the amino acid sequence of a light chain variable region as set forth in SEQ ID NO: 48

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWIS PYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDY WGQGTLVTVSS (SEQ ID NO: 48)

a human antibody or antigen-binding fragment comprising the amino acid sequence of a light chain variable region as set forth in SEQ ID NO: 49:

DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASF LYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIK (SEQ ID NO: 49)

or a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region presented in SEQ ID NO: 48 and the amino acid sequence of the light chain variable region presented in SEQ ID NO: 49.

In various embodiments of the invention are antibodies comprising a heavy chain, a light chain, or both a heavy chain and a light chain; a heavy chain variable region, a light chain variable region, or both a heavy chain variable region and a light chain variable region; wherein the heavy chain, light chain, heavy chain variable region, or light chain variable region comprises a sequence that has at least 75%, at least about 80%, at least about 85%, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least about 99% identity to the amino acid sequences set forth in SEQ ID NO: 46, 47, 48, or 49; wherein the antibody specifically binds to human PD-L1.

In various embodiments of the present invention, the anti-PD-L1 antibody is an anti-PD-L1 antibody that is a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 50:

EVQLVESGGGLVQPGGSLRLSCAASGTFFSRYWMSWVRQAPGKGLEWVANI KQDGS EKYYVDS VKGRFTIS RDN AKNS LYLQMNS LR AEDT AVYYCAREGGWFGELAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 50)

a human antibody or antigen-binding fragment comprising the light chain amino acid sequence shown in SEQ ID NO: 51:

EI VLTQS PGTLS LS PGERATLSCR ASQR VS S S YLAW YQQKPGQ APRLLIYD AS S RATGIPDRFSGSGGTDFTLTISRLEPEDFAVYYCQQYGSLPWTFGQGTKVEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KD S T YS LS S TLTLS K AD YEKHK V Y ACE VTHQGLS S P VTK S FNRGEC (SEQ ID NO: 51)

or a human antibody or antigen-binding fragment comprising the heavy chain amino acid sequence shown in SEQ ID NO: 50 and the light chain amino acid sequence shown in SEQ ID NO: 51;

a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region as set forth in SEQ ID NO: 52:

EVQLVESGGGLVQPGGSLRLSCAASGTFFSRYWMSWVRQAPGKGLEWVANI KQDGS EK Y Y VDS VKGRFTIS RDN AKNS LYLQMNS LR AEDT AV Y YC AREGGWFGEL AFDYWGQGTLVTVSS (SEQ ID NO: 52)

a human antibody or antigen-binding fragment comprising the amino acid sequence of a light chain variable region as set forth in SEQ ID NO: 53:

EI VLTQS PGTLS LS PGERATLSCR ASQR VS S S YLAW YQQKPGQ APRLLIYD AS S RATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSLPWTFGQGTKVEIK (SEQ ID NO: 53)

or a human antibody or antigen-binding fragment comprising the amino acid sequence of the heavy chain variable region as set forth in SEQ ID NO: 52 and the amino acid sequence of the light chain variable region as set forth in SEQ ID NO: 53.

In various embodiments of the invention, antibodies are provided that comprise a heavy chain, a light chain, or both a heavy chain and a light chain; a heavy chain variable region, a light chain variable region, or both a heavy chain variable region and a light chain variable region; wherein the heavy chain, light chain, heavy chain variable region, or light chain variable region comprises a sequence that has at least 75%, at least about 80%, at least about 85%, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least about 99% identity to the amino acid sequences set forth in SEQ ID NO: 50, 51, 52, or 53; wherein the antibody specifically binds to human PD-L1.

Multispecific antagonist molecules

In one aspect of the present invention, novel polypeptide-based bifunctional antagonist molecules are provided, specifically designed to simultaneously effectively neutralize the activin signaling pathway and the TGF-β signaling pathway, and comprising a first antigen-binding molecule that specifically binds to an activin ligand, and a second antigen-binding molecule that specifically binds to a TGF-β ligand. In various embodiments, the multispecific antagonist molecule comprises an isolated antibody or antigen-binding fragment thereof that specifically binds to activin, and an isolated antibody or antigen-binding fragment thereof that specifically binds to a TGF-β ligand. Importantly, these multispecific antagonists also provide favorable properties such as productivity, stability, binding affinity, biological activity, specific targeting of certain cells, targeting efficiency, and reduced toxicity.

Examples of multispecific antagonist molecules

In various embodiments, the bifunctional antagonist molecules of the present invention are selected from the group of molecules designed and comprising fusion partners presented in Table 5:

In various embodiments, a multispecific antagonist molecule according to the present invention, specifically designed to simultaneously effectively neutralize the activin signaling pathway and the TGF-β signaling pathway, and comprising a first antigen-binding molecule that specifically binds to an activin ligand and a second antigen-binding molecule that specifically binds to a TGF-β ligand, is selected from the group of molecules described in Table 6:

Bifunctional Multispecific Antagonist Molecules In another aspect of the present invention, novel polypeptide-based bifunctional multispecific antagonist molecules are provided that are specifically designed to simultaneously inhibit TGF-β, activin, and a T cell immune checkpoint (including PD1, PDL1, or CTL4). In various embodiments, the bifunctional multispecific antagonist molecule is a bifunctional multispecific molecule comprising a first antigen-binding molecule that specifically binds to an activin ligand or activin-related ligand (an "activin-binding polypeptide"), a second antigen-binding molecule that specifically binds to a TGF-β ligand (a "TGF-β-binding polypeptide"), and a third antigen-binding molecule that specifically binds to a PD-1 ligand, a PD-L1 ligand, or a CTLA-4 ligand (a "PD-1/PD-L1/CTLA-4-binding polypeptide"). Importantly, these bifunctional multispecific antagonists also provide favorable properties such as productivity, stability, binding affinity, biological activity, specific targeting of certain cells, targeting efficiency, and reduced toxicity.

Examples of bifunctional multispecific antagonist molecules In various embodiments, the bifunctional multispecific antagonist molecules of the present invention are selected from the group of molecules designed and containing fusion partners described in Tables 7 and 8:

Linkers

In various embodiments, a first antigen-binding molecule that specifically binds to activin or an activin-related ligand is attached to a second antigen-binding molecule that specifically binds to a TGF-β ligand via a linker and/or a hinge linker peptide. The linker or hinge linker may be an artificial sequence of 5, 10, 15, 20, 30, 40, or more than 40 amino acids that is relatively devoid of secondary structure or exhibits an α-helical conformation.

A peptide linker provides a covalent bond and additional structural and/or spatial flexibility between protein domains. As is known in the art, peptide linkers contain flexible amino acid residues such as glycine and serine. In various embodiments, a peptide linker may include 1-100 amino acids. GGGSGGGS (SEQ ID NO: 131). In other embodiments, the linker may comprise a GGGGS motif (SEQ ID NO: 134) _n , where n is an integer from 1 to 10. In other embodiments, the linker may also comprise amino acids other than glycine and serine. In another embodiment, the linker may comprise other protein motifs, including, but not limited to, sequences of an α-helical conformation, such as AEAAAKEAAKEAAAAKA (SEQ ID NO: 129). In various embodiments, the length and composition of the linker can be adjusted to optimize activity or performance, including, but not limited to, expression level and aggregation propensity. In another embodiment, the peptide linker can be a simple chemical bond, such as an amide bond (e.g., by chemical conjugation with PEG).

Examples of peptide linkers are shown in Table 9:

Polynucleotides

Another aspect of the present invention provides isolated nucleic acid molecules comprising a polynucleotide encoding a multispecific or bifunctional antagonist molecule as described herein. The nucleic acids in question may be single-stranded or double-stranded. Such nucleic acids may be DNA or RNA molecules. DNA includes, for example, cDNA, genomic DNA, synthetic DNA, PCR-amplified DNA, and combinations thereof. Genomic DNA encoding bifunctional antagonist molecules is obtained from genomic libraries that are available for a variety of species. Synthetic DNA is available through chemical synthesis of overlapping oligonucleotide fragments, followed by assembly of the fragments to restore part or all of the coding regions and flanking sequences. RNA can be obtained from prokaryotic expression vectors that direct high-level mRNA synthesis, such as vectors utilizing T7 promoters and RNA polymerase. cDNA is obtained from libraries prepared from mRNA isolated from various tissues that express a multispecific or bifunctional antagonist molecule. DNA molecules according to this disclosure include full-length genes, as well as polynucleotides and fragments thereof. A full-length gene may also include sequences encoding an N-terminal signal sequence.

In various embodiments, the isolated nucleic acid molecules comprise the polynucleotides described herein and further comprise a polynucleotide encoding at least one heterologous protein described herein. In various embodiments, the nucleic acid molecules further comprise polynucleotides encoding linkers or hinge linkers described herein.

In various embodiments, the recombinant nucleic acids described herein may be operably linked to one or more regulatory nucleotide sequences when expressing a multispecific or bifunctional multispecific antagonist molecule. Accordingly, the term regulatory sequence includes promoters, enhancers, and other expression control elements. Examples of regulatory sequences are described in Goeddel; Gene Expression Technology: Cnoco6bi in Enzymology, Academic Press, San Diego, Calif. (1990). Typically, the one or more regulatory nucleotide sequences may include, but are not limited to, promoter sequences, leader or signal sequences, ribosome binding sites, transcription start and stop sequences, translation start and stop sequences, and enhancer or activator sequences. Constitutive or inducible promoters known in the art are contemplated in the present invention. Promoters can be either natural promoters or hybrid promoters that combine elements of more than one promoter. The expression construct can be present in the cell on an episome, such as a plasmid, or the expression construct can be inserted into the chromosome. In various embodiments, the expression vector contains a selectable marker gene, allowing for selection of transformed host cells. Selectable marker genes are well known in the art and will vary depending on the host cell used.

In another aspect of the present invention, the nucleic acid in question is contained in an expression vector comprising a nucleotide sequence encoding a multispecific or bifunctional multispecific antagonist molecule and operably linked to at least one regulatory sequence. The term "expression vector" refers to a plasmid, phage, virus, or vector for expressing a polypeptide from a polynucleotide sequence. Vectors suitable for expression in host cells are readily available, and nucleic acid molecules are inserted into vectors using standard recombinant DNA techniques. Such vectors may include a wide range of expression control sequences that control the expression of the DNA sequence when operably linked to it and can be used in these vectors to express DNA sequences encoding a multispecific or bifunctional multispecific antagonist molecule. Such sequences suitable for controlling expression include, for example, the early and late promoters of SV40, the tet promoter, the adenovirus or immediate-early promoter of cytomegalovirus, the RSV promoters, the lac system, the trp system, the TAC or TRC system, the T7 promoter, the expression of which is directed by T7 RNA polymerase, the basic operator and promoter regions of the lambda phage, the control regions of the coat protein fd, the promoter of 3-phosphoglycerate kinase or other glycolytic enzymes, acid phosphatase promoters such as PhoS, promoters of yeast mating factors, the polyhedral promoter of the baculovirus system and other sequences known to control the expression of genes of prokaryotic or eukaryotic cells or their viruses and various combinations thereof. Clearly, the design of an expression vector can depend on factors such as the choice of host cell to be transformed and/or the type of protein to be expressed. Furthermore, the vector copy number, the ability to control this copy number, and the expression of any other proteins encoded by the vector, such as antibiotic susceptibility markers, must also be considered.

A recombinant nucleic acid according to the present invention can be obtained by ligating a cloned gene or a fragment thereof into a vector suitable for expression in either prokaryotic cells, eukaryotic cells (yeast, birds, insects, or mammals), or both. Expression vehicles for producing a recombinant multispecific or bifunctional multispecific antagonist molecule include plasmids and other vectors. For example, suitable vectors include plasmids of the following types: pBR322-derived plasmids, pEMBL-derived plasmids, pEX-derived plasmids, pBTac-derived plasmids, and pUC-derived plasmids for expression in prokaryotic cells such as E. coli.

Some mammalian expression vectors contain both prokaryotic sequences that facilitate vector propagation in bacteria and one or more eukaryotic transcription units that are expressed in eukaryotic cells. Vectors derived from pcDNAI/amp, pcDNAI/neo, pRc/CMV, pSV2gpt, pSV2neo, pSV2-dhfr, pTk2, pRSVneo, pMSG, pSVT7, pko-neo, and pHyg are examples of mammalian expression vectors suitable for transfection of eukaryotic cells. Some of these vectors are modified with sequences from bacterial plasmids, such as pBR322, to facilitate replication and selection for drug resistance in both prokaryotic and eukaryotic cells. Alternatively, derivatives of viruses such as bovine papillomavirus (BPV-1) or Epstein-Barr virus (pHEBo, a derivative of pREP, and p205) can be used for transient expression of proteins in eukaryotic cells. Examples of other viral (including retroviral) expression systems can be found below in the description of delivery systems for gene therapy. The various methods used in producing plasmids and transforming host organisms are well known in the art. For other suitable expression systems for both prokaryotic and eukaryotic cells, as well as general recombinant procedures, see Molecular Cloning: A Laboratory Manual, 2nd Ed., ed. by Sambrook, Fritsch and Maniatis (Cold Spring Harbor Laboratory Press, 1989), Chapters 16 and 17. In some cases, it may be desirable to express recombinant polypeptides using a baculovirus expression system. Examples of such baculovirus expression systems include pVL-derived vectors (such as pVL1392, pVL1393, and pVL941), pAcUW-derived vectors (such as pAcUWl), and pBlueBac-derived vectors (such as the B-gal containing pBlueBac III).

In various embodiments, a vector such as the Pcmv-Script vector (Stratagene, La Jolla, Calif.), pcDNA4 vectors (Invitrogen, Carlsbad, Calif.), and pCI-neo vectors (Promega, Madison, Wis.) will be constructed for the production of the subject multispecific or bifunctional multispecific antagonist molecule in CHO cells. As will be understood, the subject gene constructs can be used to cause expression of the subject multispecific or bifunctional multispecific antagonist molecule in cells propagated in culture, for example, to obtain proteins including fusion proteins or protein variants for purification.

Accordingly, the present disclosure further provides methods for producing the subject multispecific or bifunctional multispecific antagonist molecules. For example, a host cell transfected with an expression vector encoding the multispecific or bifunctional multispecific antagonist molecules can be cultured under suitable conditions to allow expression of the bifunctional antagonist molecule. The multispecific or bifunctional multispecific antagonist molecule can be secreted and isolated from a mixture of cells and medium containing the multispecific or bifunctional multispecific antagonist molecule. Alternatively, the multispecific or bifunctional multispecific antagonist molecule can be retained in the cytoplasm or in a membrane fraction, and the cells are collected, lysed, and the protein is isolated. Cell culture includes host cells, media, and other byproducts. Suitable cell culture media are well known in the art.

Polypeptides and proteins according to the present disclosure can be purified according to protein purification methods well known to those skilled in the art. These methods include, at one level, crude fractionation of protein and non-protein fractions. Having separated peptide polypeptides from other proteins, the peptide or polypeptide of interest can be further purified using chromatographic and electrophoretic methods to achieve partial or complete purification (or purification to homogeneity). The term "isolated polypeptide" or "purified polypeptide" as used herein is intended to refer to a composition separated from other components, wherein the polypeptide is purified to any degree relative to its natural state. Thus, a purified polypeptide also refers to a polypeptide that is free from the environment in which it may be naturally found. Typically, the term "purified" refers to a polypeptide composition that has been fractionated to remove various other components, and this composition substantially retains its expressed biological activity. When the term "substantially purified" is used, the designation will refer to a peptide or polypeptide composition in which the polypeptide or peptide forms a major component of the composition, such as comprising about 50%, about 60%, about 70%, about 80%, about 85%, or about 90% or more of the proteins in the composition.

Various methods suitable for use in purification will be well known to those skilled in the art. These include, for example, precipitation with ammonium sulfate, PEG, antibodies (immunoprecipitation), and the like, or by thermal denaturation followed by centrifugation; chromatography such as affinity chromatography (protein A columns), ion exchange, gel filtration, reverse phase, hydroxyapatite, hydrophobic interaction chromatography; isoelectric focusing; gel electrophoresis; and combinations of these methods. In accordance with common knowledge in the art, it is contemplated that the order of the various purification steps may be changed, or that certain steps may be omitted, but still provide a suitable method for obtaining a substantially purified polypeptide. Pharmaceutical compositions

In another aspect of the present disclosure, there is provided a pharmaceutical composition comprising isolated multispecific or bifunctional multispecific antagonist molecules in admixture with a pharmaceutically acceptable carrier. Such pharmaceutically acceptable carriers are well known and understood by those skilled in the art and have been described in detail (see, for example, Remington's Pharmaceutical Sciences, 18th Edition, A. R. Gennaro, ed., Mack Publishing Company, 1990). Pharmaceutically acceptable carriers can be included for the purpose of modifying, maintaining, or preserving, for example, the pH, osmolarity, viscosity, clarity, color, isotonicity, odor, sterility, stability, dissolution or release rate, adsorption, or permeability of the composition. Such pharmaceutical compositions can affect the physical state, stability, in vivo release rate, and in vivo elimination rate of the polypeptide. Suitable pharmaceutically acceptable carriers include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine, or lysine); antimicrobial agents; antioxidants (such as ascorbic acid, sodium sulfite, or sodium hydrogen sulfite); buffers (such as borate, bicarbonate, Tris-HCl, citrates, phosphates, other organic acids); bulking agents (such as mannitol or glycine); chelating agents (such as ethylenediaminetetraacetic acid (EDTA)); complexing agents (such as caffeine, polyvinylpyrrolidone, beta-cyclodextrin, or hydroxypropyl-beta-cyclodextrin); fillers; monosaccharides; disaccharides; and other carbohydrates (such as glucose, mannose, or dextrins); proteins (such as serum albumin, gelatin, or immunoglobulins); colours, flavourings and diluents; emulsifiers; hydrophilic polymers (such as polyvinylpyrrolidone); low molecular weight polypeptides; salt-forming counterions (such as sodium); preservatives (such as benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid or hydrogen peroxide); solvents (such as glycerol, propylene glycol or polyethyleneglycol); sugar alcohols (such as mannitol or sorbitol); suspending agents; surfactants or humectants (such as pluronics, PEG, sorbitan esters, polysorbates such as polysorbate 20, polysorbate 80, triton, tromethamine, lecithin, cholesterol, tyloxapal); stabilizing agents (sucrose or sorbitol); agents for increasing tonicity (such as alkali metal halides, preferably sodium or potassium chloride, mannitol, sorbitol); carriers; diluents; excipients and/or pharmaceutical adjuvants.

The primary diluent or carrier in a pharmaceutical composition may be either aqueous or non-aqueous in nature. For example, a suitable diluent or carrier may be water for injection, physiological saline, or artificial cerebrospinal fluid, optionally supplemented with other substances frequently used in parenteral compositions. Neutral buffered saline or saline mixed with serum albumin are additional typical carrier media. Other typical pharmaceutical compositions comprise a Tris buffer of approximately pH 7.0-8.5 or an acetate buffer of approximately pH 4.0-5.5, which may further include sorbitol or a suitable substitute thereof. In one embodiment of the present disclosure, the composition may be prepared for storage by mixing the selected composition, having the required degree of purity, with the optimal agents of the composition (Remington's Pharmaceutical Sciences, supra) in the form of a lyophilized mass or an aqueous solution. Additionally, the therapeutic composition can be prepared as a lyophilisate using suitable excipients, such as sucrose. The optimal pharmaceutical composition will be determined by a person skilled in the art depending on, for example, the intended route of administration, delivery format, and desired dosage.

If parenteral administration is contemplated, the therapeutic pharmaceutical compositions may be in the form of a pyrogen-free, parenterally acceptable aqueous solution containing the desired bifunctional antagonist molecule in a pharmaceutically acceptable diluent. The most suitable diluent for parenteral injection is sterile distilled water, in which the polypeptide is prepared as a sterile, isotonic solution that is properly maintained. In various embodiments, pharmaceutical compositions suitable for injection may be prepared in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiologically buffered saline. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Additionally, suspensions of the active compounds may be prepared as suitable oily injection suspensions. It is possible that the suspension may also contain suitable stabilizers or agents that increase the solubility of the compounds and allow the preparation of highly concentrated solutions.

In various embodiments, therapeutic pharmaceutical compositions can be formulated for targeted delivery using a colloidal dispersion system. Colloidal dispersion systems include macromolecular complexes, nanocapsules, microspheres, beads, and lipid-based systems, including oil-in-water emulsions, micelles, mixed micelles, and liposomes. Examples of lipids suitable for forming liposomes include phosphatidyl compounds such as phosphatidylglycerol, phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, sphingolipids, cerebrosides, and gangliosides. Exemplary phospholipids include egg phosphatidylcholine, dipalmitoylphosphatidylcholine, and distearoylphosphatidylcholine. Targeting of liposomes is also possible based on, for example, organ specificity, cell specificity and organelle specificity and is known in the art.

In various embodiments, oral administration of the pharmaceutical compositions is contemplated. Pharmaceutical compositions administered in this manner may be prepared with or without carriers commonly used in the preparation of solid dosage forms such as tablets or capsules. In solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules, etc.), one or more therapeutic compounds according to the description of the present invention may be mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or any of the following materials: (1) fillers or inert fillers, such as starches, lactose, sucrose, glucose, mannitol and/or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and/or gum arabic; (3) wetting agents, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; (5) dissolution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) humectants, such as, for example, cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof; and (10) coloring agents. In the case of capsules, tablets and pills, the pharmaceutical compositions may additionally contain buffering agents. In addition, solid compositions of a similar type can be used as fillers in soft and hard gelatin capsules, using such excipients as lactose or milk sugar, as well as high molecular weight polyethylene glycols, etc. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, peanut, corn, wheat germ, olive, castor and sesame oils), glycerin, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions also include adjuvants such as wetting agents, emulsifying and suspending agents, sweeteners, flavoring agents, coloring agents, aromatic substances and preservatives.

Various embodiments contemplate topical administration of pharmaceutical compositions to either the skin or mucous membranes. Compositions for topical administration may further include one or more of a wide variety of agents that are effective in enhancing penetration into the skin or stratum corneum. Examples of such agents include 2-pyrrolidone, N-methyl-2-pyrrolidone, dimethylacetamide, dimethylformamide, propylene glycol, methyl or isopropyl alcohol, dimethyl sulfoxide, and azone. Further, additional agents may be included to make the composition cosmetically acceptable. Examples of such agents include fats, waxes, oils, dyes, fragrances, preservatives, stabilizers, and surfactants. Keratolytic agents known in the art may also be included. Examples include salicylic acid and sulfur. Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compound can be mixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives, buffering agents, or propellants that may be necessary. Ointments, pastes, creams, and gels, in addition to the compound in question as described (e.g., in addition to the bifunctional antagonist molecule), may contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc, and zinc oxide, or mixtures thereof.

Additional pharmaceutical compositions contemplated for use herein include those comprising polypeptides in sustained- or controlled-delivery formulations. Methods for preparing various other means for sustained- or controlled-delivery, such as liposomal carriers, biodegradable microparticles, porous beads, and depot injections, are also known to those skilled in the art.

The effective amount of a therapeutically administered pharmaceutical composition will depend, for example, on the circumstances and goals of treatment. Those skilled in the art will appreciate that appropriate dosage levels for treatment will vary, in particular, depending on the molecule being delivered, the indication for which the polypeptide is used, the route of administration, the parameters (body weight, body surface area, or organ size), and the condition (age, general health) of the patient. Accordingly, the treating physician can titrate the dose and vary the route of administration to obtain the optimal therapeutic effect. A typical dose can range from about 0.1 mg/kg to about 100 mg/kg or more, depending on the aforementioned factors. Polypeptide compositions can preferably be administered by injection or intravenously. Long-acting pharmaceutical compositions can be administered every three to four days, every week, or once every two weeks, depending on the half-life and elimination rate of a particular composition. The frequency of dosing will depend on the pharmacokinetic parameters of the polypeptide in the composition used. Typically, the composition is administered until a dose that achieves the desired effect is reached. Thus, the composition may be administered as a single dose or multiple doses (at the same or different concentrations/doses) over time, or as a continuous infusion. Further determination of the appropriate dose is typically performed. Suitable doses can be selected using relevant dose-response data.

The route of administration of the pharmaceutical composition corresponds to known methods, for example, orally, by injection carried out intravenously, intraperitoneally, intracerebral (intraparenchymatous), intracerebroventricular, intramuscular, intraocular, intraarterial, intraportal or intralesional; intramedullary, intrathecal, intraventricular, transdermal, subcutaneous or intraperitoneal; as well as intranasally, enterally, locally, sublingually, urethrally, vaginally or rectally, using slow-release systems or implantable devices. If necessary, the compositions can be administered by bolus injection or continuously by infusion, or using an implantable device. Alternatively or additionally, the composition can be administered locally by implantation of a membrane, sponge or other suitable material on which the desired molecule is absorbed or encapsulated. If an implantable device is used, it can be implanted into any suitable tissue or organ, and delivery of the desired molecule will be accomplished by diffusion, delayed-release bolus, or continuous administration.

Therapeutic applications

In another aspect of the present disclosure, a method for treating or preventing various complex disease states, the pathogenesis of which involves activation of the TGF-β/activin-mediated Smad2/3 signaling pathway, is provided.

In various embodiments, the novel multispecific or bifunctional multispecific antagonist molecules of the present invention may have broad applications in the treatment of a variety of disorders including, but not limited to, the following conditions: blood disorders: ineffective erythropoiesis, anemia, pancytopenia, myelodysplastic syndromes; fibrotic diseases: non-alcoholic steatohepatitis/NASH, liver fibrosis, pulmonary fibrosis, renal fibrosis, polycystic kidney disease, cardiac fibrosis, muscle fibrosis, myelofibrosis, cutaneous fibrosis, and ocular fibrosis; hematological malignancies: multiple myeloma, leukemia, and lymphoma; Solid cancer: melanoma, multiple myeloma, lung cancer, pancreatic cancer, colorectal cancer, liver cancer, stomach cancer, kidney cancer, bladder cancer, head and neck cancer, thyroid cancer, breast cancer, ovarian cancer, endometrial cancer, testicular cancer, prostate cancer, and brain cancer; Metastasis: bone metastasis, lung metastasis, liver metastasis, brain metastasis, and sarcoma; Cancer treatment in combination with checkpoint inhibitors such as anti-PD-1, anti-PDL1, and anti-CTL4 antibodies; Neuromuscular diseases: muscular dystrophy, spinal muscular atrophy, spinal cord injury, stroke; Pain: nociceptive pain, neuropathic pain; Wasting syndromes: sarcopenia, wasting in cancer, cardiac wasting, renal wasting, rheumatoid wasting, and anorexia; Bone disorders: bone metastasis, skeletal phenomena in cancer, bone fragility, fracture, osteogenesis imperfection, and osteoporosis; Cardiovascular diseases: pulmonary hypertension, myocardial infarction, heart failure; Metabolic disorders: insulin resistance, diabetic nephropathy, chronic renal failure; Inflammatory diseases: rheumatoid arthritis, inflammatory bowel disease; Infections: SARS-CoV, cytokine storm syndrome, sepsis; and Trauma: burn injury.

The present invention provides a method for treating a cardiovascular disease in a subject, comprising administering to said subject a therapeutically effective amount (in a monotherapy regimen or in a combination therapy regimen) of a multispecific antagonist molecule according to the present invention in a pharmaceutically acceptable carrier, wherein such administration reduces inflammation and fibrosis of vascular networks and muscles, including smooth muscles, cardiac muscle and skeletal muscle. In particular, the multispecific antagonist molecule according to the present invention is useful in the treatment of heart failure, pulmonary hypertension, myocarditis, coronary heart disease, myocardial infarction, cardiac arrhythmias, heart valve disease, cardiomyopathy, pericardial disease, aortic disease, Marfan syndrome and cardiac atrophy.

The present invention provides a method for treating cardiac dysfunction or heart failure in a subject, comprising administering to the subject an effective amount of a multispecific antagonist molecule. The modulation can improve the cardiac function of said subject by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%. Improvement in cardiac function can be assessed by echocardiography to measure 1) cardiac pump function, based on ejected blood volume and ejection efficiency, and 2) myocardial function, based on myocardial contractile force.

The present invention provides a method for treating metabolic disorders in a subject, comprising administering to said subject a therapeutically effective amount (in a monotherapy regimen or in a combination therapy regimen) of a multispecific antagonist molecule according to the present invention in a pharmaceutically acceptable carrier. In particular, the multispecific antagonist molecule according to the present invention is useful in the treatment of a metabolic disease selected from obesity, dyslipidemia, diabetes mellitus, insulin resistance, sarcopenic obesity, steatosis and metabolic syndrome, as well as diabetic myopathy, nephropathy, neuropathy, retinopathy, bone loss, impaired glucose tolerance, hyperglycemia and androgen deprivation.

The present invention provides a method for treating cancer cells, as well as cancer cell metastases, in a subject, comprising administering to said subject a therapeutically effective amount (in a monotherapy regimen or in a combination therapy regimen) of a multispecific or bifunctional multispecific antagonist molecule according to the description of the present invention in a pharmaceutically acceptable carrier, wherein such administration inhibits the growth and/or proliferation of cancer cells. In particular, the multispecific or bifunctional multispecific antagonist molecule according to the present invention is useful for the treatment of disorders characterized as cancer. Such disorders include, but are not limited to, solid tumors such as cancer of the breast, respiratory tract, brain, reproductive organs, digestive tract, urinary tract, eye, liver, skin, head and neck, thyroid, parathyroid gland and their distant metastases, lymphomas, sarcomas, multiple myeloma and leukemia. Examples of breast cancer include, but are not limited to, invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in situ, and lobular carcinoma in situ. Examples of respiratory tract cancers include, but are not limited to, small cell and non-small cell lung carcinoma, as well as bronchial adenoma and pleural pulmonary blastoma. Examples of brain cancer include, but are not limited to, brainstem glioma and hypophthalmic glioma, cerebellar and cerebral astrocytoma, medullary blastoma, ependymoma, as well as neuroectodermal tumor and pineal gland tumor. Tumors of the male reproductive organs include, but are not limited to, prostate cancer and testicular cancer. Tumors of the female genital tract include, but are not limited to, endometrial cancer, cervical cancer, ovarian cancer, vaginal cancer, vulvar cancer, and uterine sarcoma. Tumors of the gastrointestinal tract include, but are not limited to, anal cancer, colon cancer, colorectal cancer, esophageal cancer, gallbladder cancer, stomach cancer, pancreatic cancer, rectal cancer, small bowel cancer, and salivary gland cancer. Tumors of the urinary tract include, but are not limited to, bladder, penile, kidney, renal pelvic, ureteral, and urethral cancers. Eye cancer includes, but is not limited to, intraocular melanoma and retinoblastoma. Examples of liver cancer include, but are not limited to, hepatocellular carcinoma (liver cell cancer with or without the fibrolamellar variant), cholangiocarcinoma (intrahepatic bile duct carcinoma), and mixed hepatocellular cholangiocarcinoma. Skin cancer includes, but is not limited to, squamous cell carcinoma, Kaposi's sarcoma, malignant melanoma, Merkel cell skin cancer, and skin cancer other than melanoma. Head and neck cancer includes, but is not limited to, cancer of the nasopharynx and lip, and cancer of the oral cavity. Lymphomas include, but are not limited to, AIDS-related lymphoma, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, Hodgkin's disease, and central nervous system lymphoma. Sarcomas include, but are not limited to, soft tissue sarcoma, osteosarcoma, malignant fibrous histiocytoma, lymphosarcoma, and rhabdomyosarcoma. Leukemias include, but are not limited to, acute myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, and hairy cell leukemia. In some embodiments, the cancer is a cancer with high levels of TNF-α and TGF-β expression, such as pancreatic cancer, gastric cancer, ovarian cancer, colorectal cancer, melanoma, leukemia, lung cancer, prostate cancer, brain cancer, bladder cancer, and head and neck cancer.

The present invention provides a method for treating chronic kidney disease (CKD) in a subject, comprising administering to said subject a therapeutically effective amount (in either a monotherapy or combination therapy regimen) of a multispecific antagonist molecule according to the present invention in a pharmaceutically acceptable carrier, wherein such administration reduces the loss of kidney function and prevents muscle atrophy or suppresses kidney fibrosis. In particular, the multispecific antagonist molecule according to the present invention is useful in the treatment of CKD, including interstitial fibrosis, renal failure and renal dialysis, as well as protein-energy malnutrition (PEW) associated with CKD. The modulation can improve CKD or PEW in said subject by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 3 by 0%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%. Improvement in renal function can be assessed by measuring the protein/creatinine ratio (PCR) in urine and glomerular filtration rate (GFR). Improvement in PEW can be assessed by measuring serum albumin and inflammatory cytokine levels, protein synthesis and degradation rates, body weight, muscle mass, physical activity, and nutritional outcomes.

The present invention provides a method for treating an autoimmune disease in a subject, comprising administering to said subject a therapeutically effective amount (in a monotherapy regimen or in a combination therapy regimen) of a multispecific antagonist molecule according to the present invention in a pharmaceutically acceptable carrier. In particular, the bifunctional antagonist molecule according to the present invention is useful in the treatment of an autoimmune disorder selected from multiple sclerosis, diabetes mellitus (type 1), glomerulonephritis, malignant myasthenia, psoriasis, systemic sclerosis and systemic lupus erythematosus, polymyositis and primary biliary cirrhosis.

The present invention provides methods for treating arthritis in a subject, comprising administering to said subject a therapeutically effective amount (in a monotherapy regimen or in a combination therapy regimen) of a multispecific antagonist molecule according to the present invention in a pharmaceutically acceptable carrier. In particular, the bifunctional antagonist molecule according to the present invention is useful in the treatment of arthritis selected from rheumatoid arthritis and osteoarthritis.

The present invention provides methods for treating anorexia in a subject, comprising administering to said subject a therapeutically effective amount (in a monotherapy regimen or in a combination therapy regimen) of a multispecific antagonist molecule according to the present invention in a pharmaceutically acceptable carrier. In particular, the bifunctional antagonist molecule according to the present invention is useful in the treatment of anorexia nervosa selected from anorexia nervosa and anorexia-cachexia syndrome.

The present invention provides methods for treating a liver disease in a subject, comprising administering to said subject a therapeutically effective amount (in a monotherapy regimen or in a combination therapy regimen) of a multispecific antagonist molecule according to the present invention in a pharmaceutically acceptable carrier. In particular, the bifunctional antagonist molecule according to the present invention is useful in the treatment of a liver disease selected from non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, alcoholic fatty liver disease, liver cirrhosis, liver failure, autoimmune hepatitis, and hepatocellular carcinoma.

The present invention provides methods for transplanting an organ or tissue in a subject, comprising administering to said subject a therapeutically effective amount (in a monotherapy regimen or in a combination therapy regimen) of a multispecific antagonist molecule according to the present invention in a pharmaceutically acceptable carrier. In particular, the multispecific antagonist molecule according to the present invention is useful in the treatment of transplantation selected from heart, kidney, liver, lung, pancreas, intestine and thymus transplantations, or from bone, tendon, cornea, skin, heart valve, nerve and vein tissue transplantations.

The present invention provides methods for treating anemia in a subject, comprising administering to said subject a therapeutically effective amount (in a monotherapy regimen or in a combination therapy regimen) of a multispecific antagonist molecule according to the present invention in a pharmaceutically acceptable carrier. In various embodiments, the anemia is selected from various anemic disorders, including anemia associated with cancer, anemia induced by chemotherapy, anemia associated with chronic renal failure, iron deficiency anemia, iron and hemochromatosis, thalassemia, sickle cell anemia, aplastic anemia, myelodysplastic syndromes, pancytopenia and bone marrow failure.

The present invention provides methods for treating fibrosis in a subject, comprising administering a therapeutically effective amount of the pharmaceutical compositions according to the invention to a subject in need thereof. In one embodiment, the subject is a human subject. In various embodiments, the fibrosis is selected from pulmonary fibrosis (such as idiopathic pulmonary fibrosis and cystic fibrosis), liver fibrosis (such as non-alcoholic steatohepatitis and cirrhosis), airway fibrosis (such as asthma), cardiac fibrosis (such as myocardial infarction, diastolic dysfunction, or valvular heart disease), renal fibrosis (such as interstitial fibrosis), myelofibrosis, idiopathic retroperitoneal fibrosis, nephrogenic fibrosing dermatopathy, intestinal fibrosis in inflammatory diseases (including Crohn's disease), keloid, scleroderma, systemic sclerosis, and arthrofibrosis.

The present invention provides methods for treating pain in a subject, comprising administering a therapeutically effective amount of the pharmaceutical compositions according to the invention to the subject in need thereof. In one embodiment, the subject is a human subject. In various embodiments, the pain is selected from neuropathic pain, inflammatory pain, or pain due to cancer.

The present invention provides methods for treating a bone disease in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition according to the invention. In one embodiment, the subject is a human subject. In various embodiments, the bone disease is selected from osteomalacia, osteoporosis, osteogenesis imperfecta, fibrodysplasia ossificans progressiva, corticosteroid-induced bone loss, bone fracture, and bone metastasis.

The present invention provides a method for inhibiting the loss of muscle mass and/or muscle function in a subject, comprising administering to said subject an effective amount of a bifunctional antagonist molecule. The modulation can attenuate the loss of muscle mass and/or muscle function in said subject by at least 5%, 10%, at least 25%, at least 50%, at least 75%, or at least 90%. Inhibition of muscle mass and muscle function loss can be assessed using imaging methods and strength tests. Examples of imaging methods for assessing muscle mass include dual-energy X-ray absorptiometry (DEXA), magnetic resonance imaging (MRI), and computed tomography (CT). Examples of muscle function tests include the grip strength test, the stair climbing test, the short-term physical performance battery (SPPB) and the 6-minute walk test, as well as the maximum inspiratory pressure (MIP) and maximum expiratory pressure (MEP), which are used to measure the strength of the respiratory muscles.

A "therapeutically effective amount" or "therapeutically effective dose" refers to the amount of a therapeutic agent administered that alleviates to some extent one or more symptoms of the disorder being treated.

A therapeutically effective dose can be initially estimated for cell culture assays by determining the IC ₅₀ (mean inhibitory concentration). The dose can then be formulated in animal models to achieve a circulating plasma concentration range that includes the IC ₅₀ determined in cell culture. This information can be used to more accurately determine useful doses in humans. Plasma levels can be measured, for example, by HPLC (high-performance liquid chromatography). The precise formulation, route of administration, and dose can be selected by the individual physician based on the patient's condition.

Dosage regimens can be adjusted to achieve the optimal desired response (e.g., a therapeutic or prophylactic response). For example, a single bolus may be administered, several divided doses (multiple, repeat, or maintenance) may be administered over a period of time, and the dose may be proportionally decreased or increased according to the therapeutic situation. It is particularly advantageous to formulate parenteral compositions in unit dosage form for ease of administration and uniformity of dosage. Unit dosage form, as used herein, refers to physically discrete units suitable as unitary dosages for the mammalian subjects to be treated, each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification of unit dosage forms in accordance with the description of the present invention will be dictated primarily by the unique characteristics of the antibody and the particular therapeutic or prophylactic effect to be achieved.

Thus, one skilled in the art will appreciate, based on the disclosure provided herein, that the dose and dosage regimen are adjusted according to methods well known in the therapeutic field. That is, the maximum tolerated dose can be readily established, as can the effective amount providing a detectable therapeutic benefit to the subject, as well as the time requirements for administering each agent to provide a detectable therapeutic benefit to the subject. Accordingly, although specific doses and dosage regimens are provided herein as examples, these examples in no way limit the dose and dosage regimen that can be used for a subject in practicing the disclosure of the present invention.

It should be noted that dosage amounts may vary depending on the type and severity of the condition to be alleviated, and may include single or multiple doses. It should also be understood that for any particular subject, specific dosage regimens should be adjusted over time in accordance with the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the dosage ranges set forth herein are provided for exemplary purposes only and are not intended to limit the scope or practical implementation of the claimed composition. Furthermore, the dosage regimen for the compositions described herein may be based on a variety of factors, including the type of disease, age, weight, gender, health condition of the subject, severity of the condition, route of administration, and the particular antibody used. Thus, the dosage regimen may vary widely, but can be determined in a routine manner using standard methods. For example, doses can be adjusted based on pharmacokinetic or pharmacodynamic parameters, which may include clinical effects such as toxic effects and/or laboratory parameters. Thus, the present disclosure encompasses dose escalation for a particular subject, as determined by one skilled in the art. Determining appropriate doses and dosage regimens is well known in the art and should be readily apparent to those skilled in the art upon familiarity with the teachings disclosed herein.

An example of a non-limiting daily dose range for a therapeutically or prophylactically effective amount of multispecific or bifunctional multispecific antagonist molecules according to the description of the present invention may be from 0.001 to 100 mg/kg, from 0.001 to 90 mg/kg, from 0.001 to 80 mg/kg, from 0.001 to 70 mg/kg, from 0.001 to 60 mg/kg, from 0.001 to 50 mg/kg, from 0.001 to 40 mg/kg, from 0.001 to 30 mg/kg, from 0.001 to 20 mg/kg, from 0.001 to 10 mg/kg, from 0.001 to 5 mg/kg, from 0.001 to 4 mg/kg, from 0.001 to 3 mg/kg, from 0.001 to 2 mg/kg, from 0.001 to 1 mg/kg, from 0.010 to 50 mg/kg, from 0.010 to 40 mg/kg, from 0.010 to 30 mg/kg, from 0.010 to 20 mg/kg, from 0.010 to 10 mg/kg, from 0.010 to 5 mg/kg, from 0.010 to 4 mg/kg, from 0.010 to 3 mg/kg, from 0.010 to 2 mg/kg, from 0.010 to 1 mg/kg, from 0.1 to 50 mg/kg, from 0.1 to 40 mg/kg, from 0.1 to 30 mg/kg, from 0.1 to 20 mg/kg, from 0.1 to 10 mg/kg, from 0.1 to 5 mg/kg, 0.1 to 4 mg/kg, 0.1 to 3 mg/kg, 0.1 to 2 mg/kg, 0.1 to 1 mg/kg, 1 to 50 mg/kg, 1 to 40 mg/kg, 1 to 30 mg/kg, 1 to 20 mg/kg, 1 to 10 mg/kg, 1 to 5 mg/kg, 1 to 4 mg/kg, 1 to 3 mg/kg, 1 to 2 mg/kg, or 1 to 1 mg/kg of body weight. It should be noted that dosage values may vary depending on the type and severity of the conditions to be relieved. It should also be understood that for any particular subject, specific dosage regimens should be adjusted over time according to individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the dosage ranges set forth herein are for exemplary purposes only and are not intended to limit the scope or practical implementation of the claimed composition.

In various embodiments, the total administered dose will achieve an antibody concentration in the blood plasma in the range of, for example, from about 1 to 1000 μg/mL, from about 1 to 750 μg/mL, from about 1 to 500 μg/mL, from about 1 to 250 μg/mL, from about 10 to 1000 μg/mL, from about 10 to 750 μg/mL, from about 10 to 500 μg/mL, from about 10 to 250 μg/mL, from about 20 to 1000 μg/mL, from about 20 to 750 μg/mL, from about 20 to 500 μg/mL, from about 20 to 250 μg/mL, from about 30 to 1000 μg/mL, from about 30 to 750 mcg/ml, from about 30 to 500 mcg/ml, from about 30 to 250 mcg/ml.

The toxicity and therapeutic index of pharmaceutical compositions according to the present invention can be determined using standard pharmaceutical methods in cell cultures or experimental animals, for example, to determine the LD ₅₀ (median lethal dose) and ED ₅₀ (median effective dose). The dose ratio between the toxic and therapeutically effective doses is the therapeutic index and can be expressed as the LD ₅₀ /ED ₅₀ ratio. Compositions that exhibit high therapeutic indices are generally preferred.

The frequency of administration of the pharmaceutical composition of the multispecific or bifunctional multispecific antagonist molecule depends on the nature of the therapy and the specific disease being treated. The subject may be treated at regular intervals, such as weekly or monthly, until the desired therapeutic result is achieved. Examples of dosage frequencies include, without limitation, once a week without interruption; once a week, every other week; once every 2 weeks; once every 3 weeks; once a week without interruption for 2 weeks, then monthly; once a week without interruption for 3 weeks, then monthly; monthly; every other month; once every three months; once every four months; once every five months; or once every six months, or once a year. Combination therapy

The terms "co-administration," "co-administered," and "in combination with," as used herein, referring to a multispecific or bifunctional multispecific antagonist molecule as described herein and one or more other therapeutic agents, are intended to mean and refer to and include the following: the simultaneous administration of such a combination of a multispecific or bifunctional multispecific antagonist molecule as described herein and a therapeutic agent(s) to a subject in need of treatment, when such components are formulated together into a unit dosage form that releases said components at substantially the same time in the body of said subject; substantially simultaneous administration of such a combination of a multispecific or bifunctional multispecific antagonist molecule according to the description of the present invention and a therapeutic agent(s) to a subject in need of treatment, wherein such components are formulated separately from one another in separate dosage forms that are taken at substantially the same time by said subject, wherein said components are subsequently released at substantially the same time in the body of said subject; and sequential administration of such a combination of a multispecific or bifunctional multispecific antagonist molecule according to the description of the present invention and a therapeutic agent(s) to a subject in need of treatment, wherein such components are formulated separately from one another in separate dosage forms that are taken by said subject sequentially with a significant time interval between each administration, wherein said components are subsequently released at substantially different times in the body of said subject; and sequentially administering such a combination of a multispecific or bifunctional multispecific antagonist molecule according to the description of the present invention and a therapeutic agent(s) to a subject in need of treatment, when such components are prepared in a unit dosage form that releases said components in a controlled manner, after which they are simultaneously, sequentially and/or partially released at the same and/or different times in the body of said subject, and wherein each part can be administered either through the same or through a different route.

In another aspect, the present disclosure provides a method of treating cancer or cancer metastasis in a subject, comprising administering a therapeutically effective amount of the pharmaceutical compositions of the invention in combination with a second therapy selected from the group consisting of: cytotoxic chemotherapy, immunotherapy, small molecule kinase targeted therapy, surgery, radiation therapy, and stem cell transplantation. In various embodiments, the combination therapy may comprise administering to the subject a therapeutically effective amount of immunotherapy, including, but not limited to, treatment using depleting antibodies against specific tumor antigens; treatment using antibody-drug conjugates; treatment with agonist, antagonist, or blocking antibodies against costimulatory or coinhibitory molecules (immune checkpoints) such as CTLA-4, PD-1, PD-L1, OX-40, CD137, TIGIT, GITR, LAG3, TIM-3, CD47, SIRPα, ICOS, and VISTA; treatment with bispecific T-cell engager activator antibodies (BiTE®) such as blinatumomab; treatment involving the administration of biological response modifiers such as the TNF family, IL-1, IL-4, IL-7, IL-12, IL-15, IL-17, IL-21, IL-22, GM-CSF (granulocyte-macrophage colony-stimulating factor), IFN-α, IFN-β, and IFN-γ; treatment with therapeutic vaccines such as sipuleucel-T; treatment with dendritic cell vaccines or tumor antigen-based peptide vaccines; treatment with chimeric antigen receptor (CAR) T cells; treatment with CAR-NK natural killer cells; treatment with tumor-infiltrating lymphocytes (TILs); treatment with adoptively transferred anti-tumor T cells (ex vivo dividing and/or TCR transgenic); treatment with TALL-104 cells; and treatment with immunostimulatory agents such as Toll-like receptor (TLR: TLR7, TLR8, and TLR 9) CpG agonists and imiquimod; wherein the combination therapy provides enhanced killing of tumor effector cells, i.e. Synergistic actions exist between a multispecific antagonist molecule or a bifunctional multispecific antagonist molecule and immunotherapy when administered together.

In various embodiments, combination therapy comprises the simultaneous administration of a multispecific or bifunctional multispecific antagonist molecule and a second agent composition in the same pharmaceutical composition or in a separate pharmaceutical composition. In various embodiments, the multispecific or bifunctional multispecific antagonist molecule composition and the second agent composition are administered sequentially, i.e., the multispecific or bifunctional multispecific antagonist molecule composition is administered before or after the second agent composition.

In various embodiments, the administration of the composition of the multispecific or bifunctional multispecific antagonist molecule and the composition of the second agent are simultaneous, i.e., the period of administration of the composition of the multispecific or bifunctional multispecific antagonist molecule and the composition of the second agent overlap with each other.

In various embodiments, the administration of the multispecific or bifunctional multispecific antagonist molecule composition and the second agent composition are not simultaneous. For example, in various embodiments, the administration of the multispecific or bifunctional multispecific antagonist molecule composition is discontinued before the administration of the second agent composition. In various embodiments, the administration of the second agent composition is discontinued before the administration of the bifunctional antagonist molecule composition.

Multispecific and/or bifunctional multispecific antagonist molecules according to the present invention can be produced using recombinant DNA techniques well known to those skilled in the art. This example generally describes the production of bifunctional antagonist molecules.

cDNAs encoding polypeptides of various novel multispecific or bifunctional multispecific antagonists were obtained by gene synthesis and subcloned into mammalian expression plasmids. CHO cells were transiently or stably transfected with mammalian expression plasmids encoding individual multispecific or bifunctional multispecific polypeptide antagonists. Transiently transfected CHO cells or stably transfected CHO pools were grown in high-density suspension cultures in a _CO2 shaking incubator at 32°C for 6-8 days. Culture medium was collected after passing through a 0.22 μm filter (Millipore Corporation, MA). Recombinantly expressed multispecific or bifunctional multispecific polypeptides were purified from culture media by protein A affinity chromatography using the AKTA PFLC system (GE Healthcare).

Example 2

Binding activities of individual multispecific or bifunctional multispecific antagonists to human ligands or target proteins were measured by biolayer interferometry (BLI) using an Octet RED96 (ForteBIO, Pall Corporation, USA). Binding analysis was performed by first capturing polypeptide multispecific or bifunctional multispecific antagonists with biosensors. To measure the rates of association and dissociation, biosensors with captured polypeptide antagonists were immersed in wells containing different concentrations of specific ligands or target proteins (such as activin A, activin B, activin AB) in kinetic buffer for 10 min followed by 10-20 min in 1x kinetic buffer. Biosensors with captured polypeptide antagonists were also immersed in wells containing 1x kinetic buffer to allow for single control subtraction to compensate for the natural dissociation of captured bifunctional antagonists. Capture sensorgrams were collected using an 8-channel detection method on the biosensor. Data were acquired and analyzed using data acquisition software v11.1 (ForteBIO, Pall Corporation, USA).

The binding affinities of the multispecific antagonist molecules A115, A116, A117, and A118 were tested using the B12 assay. As shown in Table 10, the multispecific polypeptide antagonists A115, A116, A117, and A118 were able to selectively bind to a subgroup of the TGF-β superfamily, including activin A, activin B, activin AB, GDF-8, GDF-11, TGF-β1, and TGF-β3, with high affinity. A216, a multispecific polypeptide antagonist designed in a configuration different from A116, was tested for binding to its ligand, and the data demonstrate that A216 had high binding affinities similar to A116. Thus, multispecific polypeptide antagonists designed in different configurations can achieve high ligand binding affinity.

In addition, the ligand binding affinities of the antibody-based multispecific antagonists A155 and A167 were tested using the BLI assay. As shown in Table 11, A155 and A167 are capable of binding to activin A, TGF-β1, and TGF-β3 with high affinity.

Example 3

Analysis of the Smad2/3 signaling pathway. A cell line engineered with the luciferase reporter C2C12-CAGA-luc, capable of detecting the Smad2/3 signaling pathway, was used to measure ligand signaling pathway activity in cell cultures. To measure the neutralizing activity of the multispecific antagonist, 2 nM human activin A, activin B, activin AB, myostatin, GDF-11, TGF-β1, or TGF-β3 were preincubated with increasing concentrations of each individual antagonist at 0.00004 nM, 0.0004 nM, 0.004 nM, 0.04 nM, 0.4 nM, 4 nM, 40 nM, and 400 nM for 1 hour at room temperature, and the preincubated reaction mixtures were added to C2C12-CAGA-luc reporter cell cultures. After 5 hours of incubation in a _CO2 incubator at 37°C, luciferase activity in C2C12-CAGA-luc reporter cultures was measured using a LuminoSkan Ascent (Thermo Scientific). _IC50 values were analyzed and plotted using Prism software (GraphPad Software).

As shown in Figs. 6-8, the bifunctional multispecific antagonists A115, A116, A117, and A118 A-119 and A-120 have a strong neutralizing effect on activin A and activin B (Fig. 6), TGF-β1 and TGF-β3 (Fig. 7), and GDF-8 and GDF-11 (Fig. 8) in cellular assays.

Example 4

Elevated levels of activin and TGF-β are involved in cell migration and metastasis in cancer. Both activin and TGF-β are highly upregulated in the tumor microenvironment, and the parallel upregulation of activin and TGF-β in the tumor microenvironment has been extensively documented. To mimic the parallel upregulation of activin and TGF-β1 in the tumor microenvironment, activin and TGF-β1 were added together to A549 lung cancer cell cultures. Under these conditions, the effect of A116, a multispecific activin and TGF-β antagonist, on cancer cell migration was tested in comparison with ActRTIA-Fc (an activin inhibitor) and TGFRII-Fc (a TGF-β inhibitor), respectively, in a cell migration assay.

Cancer cell migration assay. To test the effect of a multispecific activin and TGF-β antagonist on cancer cell migration, a monolayer scratch assay was performed using human non-small cell lung cancer A549 cells. A549 cells were seeded in multiwell plates and grown as monolayer cultures in DMEM (Dulbecco's modified Eagle's medium) supplemented with 10% fetal bovine serum (FBS) in a _CO2 incubator at 37°C. When the cells reached 90% confluence, the cultures were scratched with a 200 µl pipette tip to create a wound slit in the cell monolayer across each well. The cultures were then transferred to DMEM supplemented with 0.2% FBS with or without the following agents: 1) none (control), 2) activin A (50 ng/ml) + TGF- pi (5 ng/ml), 3) activin A (50 ng/ml) + TGF-β1 (5 ng/ml) + ActRTIA-Fc (2.5 μg/ml), 4) activin A (50 ng/ml) + TGF-β1 (5 ng/ml) + TGFRII-Fc (2.5 μg/ml), and 5) activin A (50 ng/ml) + TGF-β1 (5 ng/ml) + A116 (2.5 μg/ml). After 48 hours of treatment, the cell cultures were photographed using an inverted microscope equipped with a digital camera.

Fig. 10 shows the effect of the multifunctional peptide antagonist A116 on the migration of A549 lung cancer cells. As shown in panel A, the scratch gap remained large in untreated control A549 cultures after 48 hours of incubation. In contrast, with combined treatment with activin and TGF-β1, the scratch gap completely closed after 48 hours of incubation, indicating that activin and TGF-β1 significantly accelerated the migration of A549 lung cancer cells. Under the same conditions, inhibition of activin A with ActRIIA-Fc or inhibition of TGF-β1 with TGFRII-Fc did not prevent scratch gap closure within 48 hours of incubation (panels C and D), suggesting that inhibition of activin A or TGF-β1 alone was not sufficient to prevent accelerated cancer cell migration when both activin A and TGF-β1 were elevated. However, in the presence of high levels of activin and TGF-β1, addition of the multispecific antagonist A116 highly efficiently prevented scratch gap narrowing in A549 cancer cell cultures, as the scratch gap size of the A216-treated culture remained approximately the same as that of the untreated control culture (panel E). These results demonstrate that the multispecific antagonist A116 is capable of inhibiting accelerated cancer cell migration mediated in parallel by activin A and TGF-β. The results suggest that the novel multispecific activin and TGF-β antagonists disclosed in the present invention may provide a promising new approach to the treatment of cancer metastasis, as the novel antagonists can more effectively prevent cancer cell migration and metastasis by simultaneously blocking the increased levels of activin and TGF-β in the tumor microenvironment. Example 5

The literature notes that the expression levels of activin A and TGF-β are simultaneously increased in various types of malignancies, including lung cancer. To assess the effect of a multispecific activin and TGF-β antagonist on the oncogenic potential of cancer cells, a colony formation assay was performed on human non-small cell lung cancer A549 cells. To simulate the increased levels of activin and TGF-β in the cancer microenvironment, exogenous activin and TGF-β1 were added together to the cell culture medium of A549 lung cancer cell cultures. The effect of the multispecific polypeptide antagonist A116 was tested in comparison with ActRIIA-Fc and TGFFRII-Fc, respectively, in a colony formation assay.

Colony formation assay of cancer cells. To test the effect of a multispecific activin and TGF-β antagonist on the oncogenic potential of cancer cells, a colony formation assay was performed on A549 human lung cancer cell cultures. A549 lung cancer cells were seeded in 6-well plates at a density of approximately 500 cells per well in DMEM supplemented with 0.2% FBS and incubated for 14 days in the presence of the following agents: 1) activin A (50 ng/ml) + TGF-β1 (5 ng/ml), 2) activin A (50 ng/ml) + TGF-β1 (5 ng/ml) + ActRIIA-Fc (2.5 μg/ml), 3) activin A (50 ng/ml) and TGF-β1 (5 ng/ml) + TGFRII-Fc (2.5 μg/ml), and 4) activin A (50 ng/ml) and TGF-β1 (5 ng/ml) + A116 (2.5 μg/ml). Culture media were changed twice a week, and colony densities were counted on day 14, with colony formation defined as greater than 50 cells per colony. Representative microscopic images were obtained using a digital camera.

Figure 11 shows the effect of A116 on A549 colony formation in the presence of exogenously added activin A and TGF-β1. As shown in panels A-D, A549 colony densities were lower in cultures treated with ActRIIA-Fc, TGFRII-Fc, or A116, suggesting that inhibition of activin or TGF-β had an inhibitory effect on colony formation. However, the inhibitory effect of A116 on A549 colony formation was significantly greater compared to ActRIIA-Fc or TGFRII-Fc, suggesting that the parallel inhibition of activin A and TGF-β1 by A116 was more effective in suppressing colony formation by A549 cancer cells (FIG. 12). The enhanced anti-oncogenic activity of A116 suggests that the novel multispecific activin and TGF-β antagonists disclosed in this invention represent a promising new approach for the treatment of cancer.

Example 6

TGF-β is a major driver of tissue fibrosis, and the TGF-β-mediated Smad2/3 signaling pathway plays a central role in the pathogenesis of fibrosis. Like TGF-β, activin A and activin B mediate the Smad2/3 signaling pathway and play a critical role in the pathogenesis of fibrosis. TGF-β and activin A have been shown to be upregulated in tissues affected by fibrotic disease, and elevated levels of TGF-β and activin A act synergistically to promote disease progression by stimulating collagen overproduction and extracellular matrix deposition. Due to the parallel involvement of TGF-β and activin in fibrosis, dual targeting of elevated TGF-β and activin may improve treatment efficacy. Thus, to more effectively combat fibrotic diseases, it is important to investigate new antagonists capable of simultaneously inhibiting TGF-β and activin.

In this example, A116, a novel polypeptide-based multispecific antagonist capable of simultaneously neutralizing TGF-β and activin, was evaluated in comparison with ActRIIA-Fc, an activin-neutralizing protein, and TGFRII-Fc, a TGF-β-neutralizing protein, respectively, for its activity in reducing bleomycin-induced lung fibrosis in mice.

Study of a Murine Model of Bleomycin-Induced Pulmonary Fibrosis. All animal procedures were approved by the Institutional Animal Care Committee. Eight-week-old male C57BL/6 mice were purchased from Jackson Laboratories. Mice were maintained on a 12-h light-dark cycle and had ad libitum access to water and laboratory rodent chow. Mice were acclimated for 1 week before treatment. Bleomycin (Sigma) was dissolved in sterile 0.9% saline and administered as a single dose of 0.5 mg/kg per animal. Control animals received saline only. All animals received intratracheal injections by instillation (IT) of bleomycin or saline on day 0. Mice were randomly assigned to the following groups: (1) IT saline (control); (2) IT bleomycin (bleomycin), (3) IT bleomycin with ActRIIA-Fc (bleomycin+ActRIIA-Fc-Fc); (4) IT bleomycin with TGFRII-Fc (bleomycin+TGFRII-Fc); (4) IT bleomycin with A116 (bleomycin+A116). Starting from day 2, ActRIIA-Fc, TGFRII-Fc, and A-120 were administered to separate groups of mice, respectively, once a week by SC (subcutaneous) administration at a dose of 5 to 10 mg/kg, normalized to the molecular weight of each agent. After two weeks of treatment, animals were sacrificed, and right lung tissues were collected in cassettes and fixed in neutral buffered formalin. For histological evaluation, specimens were dehydrated in ethanol gradient series, cleared in xylene, and embedded in paraffin. Sections (4–6 μm thick) were cut and stained with hematoxylin and eosin (H&E), Masson's trichrome (MT), and an anti-alpha-SMA antibody coupled to HRP. Lung fibrotic injury was assessed histologically using the Ashcroft scoring system (Hubner et al. 2008. PMID: 18476815; DOI: 10.2144/000112729). Ashcroft scores were analyzed at 10x magnification.

Fig. 13 shows histological images of H&E-stained lung sections and Ashcraft scores for a mouse model of bleomycin-induced pulmonary fibrosis. The data indicate that dual inhibition of TGF-β and activin with A116 significantly reduced bleomycin-induced pulmonary fibrosis, whereas the activin inhibitor ActRIIA-Fc or the TGF-β blocker TGFRII-Fc moderately inhibited bleomycin-induced pulmonary fibrosis. Analysis of Ashcraft scores revealed that A116 treatment resulted in a greater reduction in pulmonary fibrosis compared to ActRIIA-Fc or TGFRII-Fc treatment.

Fig. 14 shows histological images of Masson trichrome-stained lung sections and a bar graph of the quantitative analysis of collagen deposition area in lung sections from mice with bleomycin-induced pulmonary fibrosis. The data demonstrate that A116 attenuated collagen deposition more effectively than ActRIIA-Fc or TGFRII-Fc in the bleomycin-induced pulmonary fibrosis model.

Fig. 15 shows histological images of immunostaining for α-SMA, a fibrosis marker, in lung sections from control and bleomycin-treated mice. Lung sections were immunostained with a primary anti-αSMA antibody in combination with an HRP-labeled secondary antibody. The results demonstrate that A116 was more effective than ActRIIA-Fc or TGFRII-Fc in preventing α-SMA induction in the lungs of bleomycin-treated mice.

Taken together, experimental data in mice with bleomycin-induced pulmonary fibrosis demonstrated that A116 is more effective than ActRIIA-Fc or TGFFRII-Fc in preventing fibrosis. The enhanced anti-fibrosis activity observed following dual inhibition of activin and TGF-β by A116 in the bleomycin-induced mouse model suggests that A116 and other novel multispecific activin and TGF-β antagonists according to the present invention may represent a more effective therapeutic approach to combating fibrotic diseases.

All articles and methods disclosed and claimed herein can be made and carried out without undue experimentation in light of the present invention. Although the articles and methods according to the present invention have been described in terms of preferred embodiments, it will be understood by those skilled in the art that changes can be made to the articles and methods without departing from the spirit and scope of the invention. All such changes and equivalents understood by those skilled in the art, whether now existing or later developed, are considered to be within the spirit and scope of the invention as defined in the claims. All patents, patent applications, and publications mentioned in the description are indicative of the levels of those skilled in the art to which the invention pertains. All patents, patent applications, and publications are herein incorporated by reference for all purposes to the same extent as if each individual publication were specifically and individually indicated to be incorporated by reference in its entirety. The invention illustratively described herein may suitably be carried out in the absence of any element(s) not specifically disclosed herein. It should therefore be understood that, although the present invention has been specifically disclosed in preferred embodiments and optional features, modification and variation of the concepts disclosed herein may be utilized by those skilled in the art, and that such modifications and variations are considered to be within the scope of the present invention as defined by the claims. Sequence listings

The nucleic acid and amino acid sequences listed in the appended sequence listing are presented using standard letter abbreviations for nucleotide bases and the three-letter code for amino acids as defined in 37 C.F.R. 1.822.

SEQ ID NOs: 1-9 are the amino acid sequences of various activin or activin-related ligands.

SEQ ID NOs: 10 and 14 are the amino acid sequences of the heavy chain of various antibodies that specifically bind to activin or an activin-related ligand.

SEQ ID NOs: 11 and 15 are the amino acid sequences of the light chain of various antibodies that specifically bind to activin or an activin-related ligand.

SEQ ID NOs: 12 and 16 are the amino acid sequences of the heavy chain variable region of various antibodies that specifically bind to activin or an activin-related ligand.

SEQ ID NOs: 13 and 17 are the amino acid sequences of the light chain variable region of various antibodies that specifically bind to activin or an activin-related ligand.

SEQ ID NOs: 18-21 are the amino acid sequences of various TGF-β ligands.

SEQ ID NO: 22 is the amino acid sequence of the heavy chain of an antibody that specifically binds to TGF-β ligand.

SEQ ID NO: 23 is the amino acid sequence of the light chain of an antibody that specifically binds to TGF-β ligand.

SEQ ID NO: 24 is the amino acid sequence of the heavy chain variable region of an antibody that specifically binds to TGF-β ligand.

SEQ ID NO: 25 is the amino acid sequence of the light chain variable region of an antibody that specifically binds to TGF-β ligand.

SEQ ID NO: 26, 30, 38, and 42 are the amino acid sequences of the heavy chain of various antibodies that specifically bind to PD-1 ligand.

SEQ ID NO: 27, 31, 39, and 43 are the amino acid sequences of the light chain of various antibodies that specifically bind to PD-1 ligand.

SEQ ID NOs: 28, 32, 40, and 44 are the amino acid sequences of the heavy chain variable region of various antibodies that specifically bind to PD-1 ligand.

SEQ ID NOs: 29, 33, 41, and 45 are the amino acid sequences of the light chain variable region of various antibodies that specifically bind to PD-1 ligand.

SEQ ID NO: 34 is the amino acid sequence of the heavy chain of an antibody that specifically binds to CTLA-4 ligand.

SEQ ID NO: 35 is the amino acid sequence of a light chain of an antibody that specifically binds to CTLA-4 ligand.

SEQ ID NO: 36 is the amino acid sequence of the heavy chain variable region of an antibody that specifically binds to CTLA-4 ligand.

SEQ ID NO: 37 is the amino acid sequence of the light chain variable region of an antibody that specifically binds to CTLA-4 ligand.

SEQ ID NOs: 47 and 51 are the amino acid sequences of the light chain of various antibodies that specifically bind to PD-L1 ligand.

SEQ ID NOs: 48 and 52 are the amino acid sequences of the heavy chain variable region of various antibodies that specifically bind to PD-L1 ligand.

SEQ ID NOs: 49 and 53 are the amino acid sequences of the light chain variable region of various antibodies that specifically bind to PD-L1 ligand.

SEQ ID NOs: 54-71 are the amino acid sequences of various multispecific antagonist molecules that specifically bind to activin or an activin-related ligand and that specifically bind to a TGF-β ligand.

SEQ ID NO: 72-84 are the amino acid sequences of the heavy chain of a multispecific antagonist molecule that specifically binds to activin or an activin-related ligand or a TGF-β ligand.

SEQ ID NO: 85-120 are the amino acid sequences of the heavy chain of a bifunctional multispecific antagonist molecule that specifically binds to activin or an activin-related ligand, a TGF-β ligand, and a PD-1 ligand or a CTLA-4 ligand or a PD-L1 ligand.

SEQ ID NO: 121 is the amino acid sequence of a multispecific antagonist molecule that specifically binds to activin or an activin-related ligand and that specifically binds to a TGF-β ligand.

SEQ ID NO: 122 is the amino acid sequence of the heavy chain of a multispecific antagonist molecule that specifically binds to activin or an activin-related ligand or a TGF-β ligand.

SEQ ID NO: 123 is the amino acid sequence of a light chain of a multispecific antagonist molecule that specifically binds to activin or an activin-related ligand or a TGF-β ligand.

SEQ ID NOs: 124-143 are the amino acid sequences of various peptide linker sequences.

SEQUENCE LISTINGS

--->

SEQUENCE LIST

<110> HAN, HQ

ZHOU, XIAOLAN

<120> NEW BIFUNCTIONAL MULTISPECIFIC ANTAGONISTS,

CAPABLE OF INHIBITING

MULTIPLE LIGANDS OF THE TGF-BETA FAMILY AND THEIR APPLICATIONS

<130> CACAG1.0009WO

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<170> PatentIn version 3.5

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Pro Gly Ser Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys

145 150 155 160

Val Thr Cys Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr

165 170 175

Leu Cys Gly Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg

180 185 190

Lys Ala Thr Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly

195 200 205

Lys Cys Ile Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly

210 215 220

Lys Lys Cys Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu

225 230 235 240

Cys Asp Glu Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala

245 250 255

Ser Asp Asn Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala

260 265 270

Cys Ser Ser Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn

275 280 285

Ser Ile Ser Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp

290 295 300

Tyr Ser Phe Pro Ile Ser Ser Ile Leu Glu Trp

305 310 315

<210> 4

<211> 315

<212> PRT

<213> Homo sapiens

<400> 4

Gly Asn Cys Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu

1 5 10 15

Tyr Lys Thr Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu

20 25 30

Ser Thr Ser Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys

35 40 45

Trp Met Ile Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu

50 55 60

Thr Cys Glu Asn Val Asp Cys Gly Pro Gly Gln Ser Cys Val Val Asp

65 70 75 80

Gln Thr Gly Ser Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile

85 90 95

Thr Trp Lys Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn

100 105 110

Glu Cys Ala Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu

115 120 125

Val Gln Tyr Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys

130 135 140

Pro Gly Ser Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys

145 150 155 160

Val Thr Cys Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr

165 170 175

Leu Cys Gly Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg

180 185 190

Lys Ala Thr Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly

195 200 205

Lys Cys Ile Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly

210 215 220

Lys Lys Cys Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu

225 230 235 240

Cys Asp Glu Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala

245 250 255

Ser Asp Asn Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala

260 265 270

Cys Ser Ser Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn

275 280 285

Ser Ile Ser Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp

290 295 300

Tyr Ser Phe Pro Ile Ser Ser Ile Leu Glu Trp

305 310 315

<210> 5

<211> 288

<212> PRT

<213> Homo sapiens

<400> 5

Gly Asn Cys Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu

1 5 10 15

Tyr Lys Thr Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu

20 25 30

Ser Thr Ser Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys

35 40 45

Trp Met Ile Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu

50 55 60

Thr Cys Glu Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn

65 70 75 80

Lys Lys Asn Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile

85 90 95

Thr Trp Lys Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn

100 105 110

Glu Cys Ala Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu

115 120 125

Val Gln Tyr Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys

130 135 140

Pro Gly Ser Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys

145 150 155 160

Val Thr Cys Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr

165 170 175

Leu Cys Gly Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg

180 185 190

Lys Ala Thr Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly

195 200 205

Lys Cys Ile Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly

210 215 220

Lys Lys Cys Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu

225 230 235 240

Cys Asp Glu Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala

245 250 255

Ser Asp Asn Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala

260 265 270

Cys Ser Ser Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn

275 280 285

<210> 6

<211> 110

<212> PRT

<213> Artificial

<220>

<223> Modified human ActRIIB ECD

<400> 6

Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg

1 5 10 15

Thr Asn Gln Ser Gly Leu Glu Arg Cys Glu Gly Asp Gln Asp Lys Arg

20 25 30

Leu His Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu

35 40 45

Val Lys Lys Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Gln

50 55 60

Glu Cys Val Ala Thr Lys Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys

65 70 75 80

Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly

85 90 95

Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr

100 105 110

<210> 7

<211> 110

<212> PRT

<213> Artificial

<220>

<223> Modified human ActRIIB ECD

<400> 7

Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg

1 5 10 15

Thr Asn Gln Ser Gly Leu Glu Arg Cys Tyr Gly Asp Lys Asp Lys Arg

20 25 30

Arg His Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu

35 40 45

Val Lys Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln

50 55 60

Glu Cys Val Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys

65 70 75 80

Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly

85 90 95

Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr

100 105 110

<210> 8

<211> 107

<212> PRT

<213> Artificial

<220>

<223> Modified human ActRIIB ECD

<400> 8

Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg

1 5 10 15

Thr Asn Gln Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg

20 25 30

Leu His Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu

35 40 45

Val Lys Lys Gly Cys Trp Asp Asp Asp Phe Asn Cys Tyr Asp Arg Gln

50 55 60

Glu Cys Val Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys

65 70 75 80

Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly

85 90 95

Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro Thr

100 105

<210> 9

<211> 109

<212> PRT

<213> Artificial

<220>

<223> Modified human ActRIIA ECD

<400> 9

Gly Ala Ile Leu Gly Arg Ser Glu Thr Gln Glu Cys Leu Phe Tyr Asn

1 5 10 15

Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln Thr Gly Val Glu Pro Cys

20 25 30

Glu Gly Glu Lys Asp Lys Arg Leu His Cys Tyr Ala Thr Trp Arg Asn

35 40 45

Ile Ser Gly Ser Ile Glu Ile Val Lys Lys Gly Cys Trp Leu Asp Asp

50 55 60

Phe Asn Cys Tyr Asp Arg Thr Asp Cys Val Glu Thr Glu Glu Asn Pro

65 70 75 80

Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn Met Cys Asn Glu Lys Phe

85 90 95

Ser Tyr Phe Pro Glu Met Glu Val Thr Gln Pro Thr Ser

100 105

<210> 10

<211> 452

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the heavy chain of the antibody

against activin

<400> 10

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Gly Leu Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ile Pro Tyr Asn Gly Asn Thr Asn Ser Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Arg Asp Arg Asp Tyr Gly Val Asn Tyr Asp Ala Phe Asp Ile Trp

100 105 110

Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

225 230 235 240

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Val Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly Lys

450

<210> 11

<211> 213

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the light chain of the antibody

against activin A

<400> 11

Ser Tyr Glu Val Thr Gln Ala Pro Ser Val Ser Val Ser Pro Gly Gln

1 5 10 15

Thr Ala Ser Ile Thr Cys Ser Gly Asp Lys Leu Gly Asp Lys Tyr Ala

20 25 30

Cys Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Val Leu Val Ile Tyr

35 40 45

Gln Asp Ser Lys Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser

50 55 60

Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Met

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Trp Asp Ser Ser Thr Ala Val

85 90 95

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Arg Thr Val Ala Ala Pro

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 12

<211> 122

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

heavy chain antibody against activin A

<400> 12

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Gly Leu Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ile Pro Tyr Asn Gly Asn Thr Asn Ser Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Arg Asp Arg Asp Tyr Gly Val Asn Tyr Asp Ala Phe Asp Ile Trp

100 105 110

Gly Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 13

<211> 106

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

light chain antibody against activin A

<400> 13

Ser Tyr Glu Val Thr Gln Ala Pro Ser Val Ser Val Ser Pro Gly Gln

1 5 10 15

Thr Ala Ser Ile Thr Cys Ser Gly Asp Lys Leu Gly Asp Lys Tyr Ala

20 25 30

Cys Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Val Leu Val Ile Tyr

35 40 45

Gln Asp Ser Lys Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser

50 55 60

Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Met

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Trp Asp Ser Ser Thr Ala Val

85 90 95

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105

<210> 14

<211> 453

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the heavy chain of the antibody

against activin A

<400> 14

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Phe Ser Ser His

20 25 30

Phe Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Leu Tyr Thr Gly Gly Thr Ser Phe Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Ser Met Ser Val Gly Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Ala Arg Ser Gly Ile Thr Phe Thr Gly Ile Ile Val Pro Gly Ser

100 105 110

Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser

115 120 125

Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr

130 135 140

Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro

145 150 155 160

Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val

165 170 175

His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser

180 185 190

Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr

195 200 205

Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val

210 215 220

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe

225 230 235 240

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

245 250 255

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

260 265 270

Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val

275 280 285

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser

290 295 300

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

305 310 315 320

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser

325 330 335

Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

340 345 350

Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu

405 410 415

Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Leu Gly Lys

450

<210> 15

<211> 215

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the light chain of the antibody

against activin A

<400> 15

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 16

<211> 126

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

heavy chain antibody against activin A

<400> 16

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Phe Ser Ser His

20 25 30

Phe Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Leu Tyr Thr Gly Gly Thr Ser Phe Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Ser Met Ser Val Gly Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Ala Arg Ser Gly Ile Thr Phe Thr Gly Ile Ile Val Pro Gly Ser

100 105 110

Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

115 120 125

<210> 17

<211> 108

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

light chain antibody against activin A

<400> 17

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 18

<211> 567

<212> PRT

<213> Homo sapiens

<400> 18

Met Gly Arg Gly Leu Leu Arg Gly Leu Trp Pro Leu His Ile Val Leu

1 5 10 15

Trp Thr Arg Ile Ala Ser Thr Ile Pro Pro His Val Gln Lys Ser Val

20 25 30

Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro

35 40 45

Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln

50 55 60

Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro

65 70 75 80

Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr

85 90 95

Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile

100 105 110

Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys

115 120 125

Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn

130 135 140

Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp Leu

145 150 155 160

Leu Leu Val Ile Phe Gln Val Thr Gly Ile Ser Leu Leu Pro Pro Leu

165 170 175

Gly Val Ala Ile Ser Val Ile Ile Ile Phe Tyr Cys Tyr Arg Val Asn

180 185 190

Arg Gln Gln Lys Leu Ser Ser Thr Trp Glu Thr Gly Lys Thr Arg Lys

195 200 205

Leu Met Glu Phe Ser Glu His Cys Ala Ile Ile Leu Glu Asp Asp Arg

210 215 220

Ser Asp Ile Ser Ser Thr Cys Ala Asn Asn Ile Asn His Asn Thr Glu

225 230 235 240

Leu Leu Pro Ile Glu Leu Asp Thr Leu Val Gly Lys Gly Arg Phe Ala

245 250 255

Glu Val Tyr Lys Ala Lys Leu Lys Gln Asn Thr Ser Glu Gln Phe Glu

260 265 270

Thr Val Ala Val Lys Ile Phe Pro Tyr Glu Glu Tyr Ala Ser Trp Lys

275 280 285

Thr Glu Lys Asp Ile Phe Ser Asp Ile Asn Leu Lys His Glu Asn Ile

290 295 300

Leu Gln Phe Leu Thr Ala Glu Glu Arg Lys Thr Glu Leu Gly Lys Gln

305 310 315 320

Tyr Trp Leu Ile Thr Ala Phe His Ala Lys Gly Asn Leu Gln Glu Tyr

325 330 335

Leu Thr Arg His Val Ile Ser Trp Glu Asp Leu Arg Lys Leu Gly Ser

340 345 350

Ser Leu Ala Arg Gly Ile Ala His Leu His Ser Asp His Thr Pro Cys

355 360 365

Gly Arg Pro Lys Met Pro Ile Val His Arg Asp Leu Lys Ser Ser Asn

370 375 380

Ile Leu Val Lys Asn Asp Leu Thr Cys Cys Leu Cys Asp Phe Gly Leu

385 390 395 400

Ser Leu Arg Leu Asp Pro Thr Leu Ser Val Asp Asp Leu Ala Asn Ser

405 410 415

Gly Gln Val Gly Thr Ala Arg Tyr Met Ala Pro Glu Val Leu Glu Ser

420 425 430

Arg Met Asn Leu Glu Asn Val Glu Ser Phe Lys Gln Thr Asp Val Tyr

435 440 445

Ser Met Ala Leu Val Leu Trp Glu Met Thr Ser Arg Cys Asn Ala Val

450 455 460

Gly Glu Val Lys Asp Tyr Glu Pro Pro Phe Gly Ser Lys Val Arg Glu

465 470 475 480

His Pro Cys Val Glu Ser Met Lys Asp Asn Val Leu Arg Asp Arg Gly

485 490 495

Arg Pro Glu Ile Pro Ser Phe Trp Leu Asn His Gln Gly Ile Gln Met

500 505 510

Val Cys Glu Thr Leu Thr Glu Cys Trp Asp His Asp Pro Glu Ala Arg

515 520 525

Leu Thr Ala Gln Cys Val Ala Glu Arg Phe Ser Glu Leu Glu His Leu

530 535 540

Asp Arg Leu Ser Gly Arg Ser Cys Ser Glu Glu Lys Ile Pro Glu Asp

545 550 555 560

Gly Ser Leu Asn Thr Thr Lys

565

<210> 19

<211> 137

<212> PRT

<213> Homo sapiens

<400> 19

Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp Met Ile Val

1 5 10 15

Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys

20 25 30

Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn

35 40 45

Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala

50 55 60

Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His

65 70 75 80

Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser

85 90 95

Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe

100 105 110

Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser

115 120 125

Glu Glu Tyr Asn Thr Ser Asn Pro Asp

130 135

<210> 20

<211> 592

<212> PRT

<213> Homo sapiens

<400> 20

Met Gly Arg Gly Leu Leu Arg Gly Leu Trp Pro Leu His Ile Val Leu

1 5 10 15

Trp Thr Arg Ile Ala Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp

20 25 30

Val Glu Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn

35 40 45

Arg Thr Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr

50 55 60

Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp

65 70 75 80

Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys

85 90 95

Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val

100 105 110

Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp

115 120 125

Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro

130 135 140

Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met

145 150 155 160

Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu

165 170 175

Glu Tyr Asn Thr Ser Asn Pro Asp Leu Leu Leu Val Ile Phe Gln Val

180 185 190

Thr Gly Ile Ser Leu Leu Pro Pro Leu Gly Val Ala Ile Ser Val Ile

195 200 205

Ile Ile Phe Tyr Cys Tyr Arg Val Asn Arg Gln Gln Lys Leu Ser Ser

210 215 220

Thr Trp Glu Thr Gly Lys Thr Arg Lys Leu Met Glu Phe Ser Glu His

225 230 235 240

Cys Ala Ile Ile Leu Glu Asp Asp Arg Ser Asp Ile Ser Ser Thr Cys

245 250 255

Ala Asn Asn Ile Asn His Asn Thr Glu Leu Leu Pro Ile Glu Leu Asp

260 265 270

Thr Leu Val Gly Lys Gly Arg Phe Ala Glu Val Tyr Lys Ala Lys Leu

275 280 285

Lys Gln Asn Thr Ser Glu Gln Phe Glu Thr Val Ala Val Lys Ile Phe

290 295 300

Pro Tyr Glu Glu Tyr Ala Ser Trp Lys Thr Glu Lys Asp Ile Phe Ser

305 310 315 320

Asp Ile Asn Leu Lys His Glu Asn Ile Leu Gln Phe Leu Thr Ala Glu

325 330 335

Glu Arg Lys Thr Glu Leu Gly Lys Gln Tyr Trp Leu Ile Thr Ala Phe

340 345 350

His Ala Lys Gly Asn Leu Gln Glu Tyr Leu Thr Arg His Val Ile Ser

355 360 365

Trp Glu Asp Leu Arg Lys Leu Gly Ser Ser Leu Ala Arg Gly Ile Ala

370 375 380

His Leu His Ser Asp His Thr Pro Cys Gly Arg Pro Lys Met Pro Ile

385 390 395 400

Val His Arg Asp Leu Lys Ser Ser Asn Ile Leu Val Lys Asn Asp Leu

405 410 415

Thr Cys Cys Leu Cys Asp Phe Gly Leu Ser Leu Arg Leu Asp Pro Thr

420 425 430

Leu Ser Val Asp Asp Leu Ala Asn Ser Gly Gln Val Gly Thr Ala Arg

435 440 445

Tyr Met Ala Pro Glu Val Leu Glu Ser Arg Met Asn Leu Glu Asn Val

450 455 460

Glu Ser Phe Lys Gln Thr Asp Val Tyr Ser Met Ala Leu Val Leu Trp

465 470 475 480

Glu Met Thr Ser Arg Cys Asn Ala Val Gly Glu Val Lys Asp Tyr Glu

485 490 495

Pro Pro Phe Gly Ser Lys Val Arg Glu His Pro Cys Val Glu Ser Met

500 505 510

Lys Asp Asn Val Leu Arg Asp Arg Gly Arg Pro Glu Ile Pro Ser Phe

515 520 525

Trp Leu Asn His Gln Gly Ile Gln Met Val Cys Glu Thr Leu Thr Glu

530 535 540

Cys Trp Asp His Asp Pro Glu Ala Arg Leu Thr Ala Gln Cys Val Ala

545 550 555 560

Glu Arg Phe Ser Glu Leu Glu His Leu Asp Arg Leu Ser Gly Arg Ser

565 570 575

Cys Ser Glu Glu Lys Ile Pro Glu Asp Gly Ser Leu Asn Thr Thr Lys

580 585 590

<210> 21

<211> 162

<212> PRT

<213> Homo sapiens

<400> 21

Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met Glu Ala Gln

1 5 10 15

Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala His Pro Leu

20 25 30

Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val

35 40 45

Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys

50 55 60

Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys

65 70 75 80

Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu

85 90 95

Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr His

100 105 110

Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu

115 120 125

Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp

130 135 140

Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn

145 150 155 160

Pro Asp

<210> 22

<211> 447

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the heavy chain of the antibody

against TGF-beta

<400> 22

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440 445

<210> 23

<211> 215

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the light chain of the antibody

against TGF-beta

<400> 23

Glu Thr Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Leu Gly Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Pro Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Ala Asp Ser Pro

85 90 95

Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 24

<211> 120

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

heavy chain anti-TGF-beta antibody

<400> 24

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 25

<211> 108

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

light chain anti-TGF-beta antibody

<400> 25

Glu Thr Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Leu Gly Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Pro Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Ala Asp Ser Pro

85 90 95

Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 26

<211> 447

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the heavy chain of the antibody

against PD-1

<400> 26

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440 445

<210> 27

<211> 218

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the light chain of the antibody

against PD-1

<400> 27

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Lys Gly Val Ser Thr Ser

20 25 30

Gly Tyr Ser Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Ala Ser Tyr Leu Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Ser Arg

85 90 95

Asp Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg

100 105 110

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

115 120 125

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

130 135 140

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

145 150 155 160

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

180 185 190

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

195 200 205

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 28

<211> 120

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

heavy chain anti-PD-1 antibody

<400> 28

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 29

<211> 111

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

light chain anti-PD-1 antibody

<400> 29

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Lys Gly Val Ser Thr Ser

20 25 30

Gly Tyr Ser Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Ala Ser Tyr Leu Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Ser Arg

85 90 95

Asp Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 30

<211> 440

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the heavy chain of the antibody

against PD-1

<400> 30

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Asp Cys Lys Ala Ser Gly Ile Thr Phe Ser Asn Ser

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Trp Tyr Asp Gly Ser Lys Arg Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Thr Asn Asp Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

100 105 110

Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser

115 120 125

Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp

130 135 140

Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr

145 150 155 160

Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr

165 170 175

Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys

180 185 190

Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp

195 200 205

Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala

210 215 220

Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

225 230 235 240

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

245 250 255

Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val

260 265 270

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

275 280 285

Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

290 295 300

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly

305 310 315 320

Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

325 330 335

Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr

340 345 350

Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser

355 360 365

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

370 375 380

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

385 390 395 400

Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe

405 410 415

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

420 425 430

Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 31

<211> 214

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the light chain of the antibody

against PD-1

<400> 31

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Ser Asn Trp Pro Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 32

<211> 113

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

heavy chain anti-PD-1 antibody

<400> 32

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Asp Cys Lys Ala Ser Gly Ile Thr Phe Ser Asn Ser

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Trp Tyr Asp Gly Ser Lys Arg Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Thr Asn Asp Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

100 105 110

Ser

<210> 33

<211> 107

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

light chain anti-PD-1 antibody

<400> 33

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Ser Asn Trp Pro Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 34

<211> 448

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the heavy chain of the antibody

against CTLA-4

<400> 34

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 35

<211> 215

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the light chain of the antibody

against CTLA-4

<400> 35

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Phe Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 36

<211> 118

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

heavy chain anti-CTLA-4 antibody

<400> 36

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 37

<211> 108

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

light chain anti-CTLA-4 antibody

<400> 37

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Phe Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 38

<211> 444

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the heavy chain of the antibody

against PD-1

<400> 38

Glu Val Gln Leu Leu Glu Ser Gly Gly Val Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Phe

20 25 30

Gly Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Gly Gly Gly Arg Asp Thr Tyr Phe Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Lys Gly Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Lys Trp Gly Asn Ile Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro

210 215 220

Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe

225 230 235 240

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

245 250 255

Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe

260 265 270

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

275 280 285

Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

290 295 300

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

305 310 315 320

Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala

325 330 335

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln

340 345 350

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

355 360 365

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

370 375 380

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser

385 390 395 400

Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu

405 410 415

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

420 425 430

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 39

<211> 214

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the light chain of the antibody

against PD-1

<400> 39

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Ser Ile Thr Ile Thr Cys Arg Ala Ser Leu Ser Ile Asn Thr Phe

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu His Gly Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Arg Thr Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ser Asn Thr Pro Phe

85 90 95

Thr Phe Gly Pro Gly Thr Val Val Asp Phe Arg Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 40

<211> 117

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

heavy chain anti-PD-1 antibody

<400> 40

Glu Val Gln Leu Leu Glu Ser Gly Gly Val Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Phe

20 25 30

Gly Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Gly Gly Gly Arg Asp Thr Tyr Phe Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Lys Gly Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Lys Trp Gly Asn Ile Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 41

<211> 107

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

light chain anti-PD-1 antibody

<400> 41

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Ser Ile Thr Ile Thr Cys Arg Ala Ser Leu Ser Ile Asn Thr Phe

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu His Gly Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Arg Thr Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ser Asn Thr Pro Phe

85 90 95

Thr Phe Gly Pro Gly Thr Val Val Asp Phe Arg

100 105

<210> 42

<211> 450

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the heavy chain of the antibody

against PD-1

<400> 42

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

405 410 415

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly Lys

450

<210> 43

<211> 216

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the light chain of the antibody

against PD-1

<400> 43

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser

85 90 95

Ser Thr Arg Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly Gln

100 105 110

Pro Lys Ala Asn Pro Thr Val Thr Leu Phe Pro Pro Ser Ser Glu Glu

115 120 125

Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr

130 135 140

Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Gly Ser Pro Val Lys

145 150 155 160

Ala Gly Val Glu Thr Thr Lys Pro Ser Lys Gln Ser Asn Asn Lys Tyr

165 170 175

Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His

180 185 190

Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys

195 200 205

Thr Val Ala Pro Thr Glu Cys Ser

210 215

<210> 44

<211> 120

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

heavy chain anti-PD-1 antibody

<400> 44

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 45

<211> 110

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

light chain anti-PD-1 antibody

<400> 45

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser

85 90 95

Ser Thr Arg Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu

100 105 110

<210> 46

<211> 448

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the heavy chain of the antibody

against PD-L1

<400> 46

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 47

<211> 214

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the light chain of the antibody

against PD-L1

<400> 47

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Ser Thr Ala

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Leu Tyr His Pro Ala

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 48

<211> 118

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

heavy chain antibody against PD-L1

<400> 48

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 49

<211> 107

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

light chain anti-PD-1 antibody

<400> 49

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Ser Thr Ala

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Leu Tyr His Pro Ala

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 50

<211> 451

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the heavy chain of the antibody

against PD-L1

<400> 50

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr

20 25 30

Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Asn Ile Lys Gln Asp Gly Ser Glu Lys Tyr Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Glu Gly Gly Trp Phe Gly Glu Leu Ala Phe Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

115 120 125

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala

130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val

145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala

165 170 175

Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val

180 185 190

Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His

195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys

210 215 220

Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly

225 230 235 240

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

245 250 255

Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His

260 265 270

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val

275 280 285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr

290 295 300

Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly

305 310 315 320

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Ser Ile

325 330 335

Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

340 345 350

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly Lys

450

<210> 51

<211> 215

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the light chain of the antibody

against PD-L1

<400> 51

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Arg Val Ser Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Asp Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Leu Pro

85 90 95

Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 52

<211> 121

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

heavy chain antibody against PD-L1

<400> 52

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr

20 25 30

Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Asn Ile Lys Gln Asp Gly Ser Glu Lys Tyr Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Glu Gly Gly Trp Phe Gly Glu Leu Ala Phe Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 53

<211> 108

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of the variable region

light chain anti-PD-1 antibody

<400> 53

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Arg Val Ser Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Asp Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Leu Pro

85 90 95

Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 54

<211> 509

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A115

<400> 54

Glu Thr Gln Glu Cys Leu Phe Phe Asn Ala Asn Trp Glu Lys Asp Arg

1 5 10 15

Thr Asn Gln Thr Gly Val Glu Pro Cys Tyr Gly Asp Lys Asp Lys Arg

20 25 30

Arg His Cys Phe Ala Thr Trp Lys Asn Ile Ser Gly Ser Ile Glu Ile

35 40 45

Val Lys Gln Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Thr

50 55 60

Asp Cys Val Glu Lys Lys Asp Ser Pro Glu Val Tyr Phe Cys Cys Cys

65 70 75 80

Glu Gly Asn Met Cys Asn Glu Lys Phe Ser Tyr Phe Pro Glu Met Glu

85 90 95

Val Thr Gln Pro Thr Ser Asn Pro Val Thr Pro Lys Pro Pro Gly Gly

100 105 110

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys

115 120 125

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu

130 135 140

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

145 150 155 160

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

165 170 175

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

180 185 190

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

195 200 205

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

210 215 220

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

225 230 235 240

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

245 250 255

Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln

260 265 270

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

275 280 285

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

290 295 300

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

305 310 315 320

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

325 330 335

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

340 345 350

Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

355 360 365

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn

370 375 380

Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu

385 390 395 400

Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser

405 410 415

Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu

420 425 430

Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu

435 440 445

Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu

450 455 460

Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly

465 470 475 480

Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn

485 490 495

Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

500 505

<210> 55

<211> 534

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A116

<400> 55

Glu Thr Gln Glu Cys Leu Phe Phe Asn Ala Asn Trp Glu Lys Asp Arg

1 5 10 15

Thr Asn Gln Thr Gly Val Glu Pro Cys Tyr Gly Asp Lys Asp Lys Arg

20 25 30

Arg His Cys Phe Ala Thr Trp Lys Asn Ile Ser Gly Ser Ile Glu Ile

35 40 45

Val Lys Gln Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Thr

50 55 60

Asp Cys Val Glu Lys Lys Asp Ser Pro Glu Val Tyr Phe Cys Cys Cys

65 70 75 80

Glu Gly Asn Met Cys Asn Glu Lys Phe Ser Tyr Phe Pro Glu Met Glu

85 90 95

Val Thr Gln Pro Thr Ser Asn Pro Val Thr Pro Lys Pro Pro Gly Gly

100 105 110

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys

115 120 125

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu

130 135 140

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

145 150 155 160

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

165 170 175

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

180 185 190

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

195 200 205

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

210 215 220

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

225 230 235 240

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

245 250 255

Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln

260 265 270

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

275 280 285

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

290 295 300

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

305 310 315 320

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

325 330 335

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

340 345 350

Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

355 360 365

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

370 375 380

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

385 390 395 400

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

405 410 415

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

420 425 430

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

435 440 445

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

450 455 460

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

465 470 475 480

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

485 490 495

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

500 505 510

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

515 520 525

Asn Thr Ser Asn Pro Asp

530

<210> 56

<211> 509

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A140

<400> 56

Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg

1 5 10 15

Thr Asn Gln Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg

20 25 30

Leu His Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu

35 40 45

Val Lys Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln

50 55 60

Glu Cys Val Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys

65 70 75 80

Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly

85 90 95

Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly

100 105 110

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys

115 120 125

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu

130 135 140

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

145 150 155 160

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

165 170 175

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

180 185 190

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

195 200 205

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

210 215 220

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

225 230 235 240

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

245 250 255

Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln

260 265 270

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

275 280 285

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

290 295 300

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

305 310 315 320

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

325 330 335

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

340 345 350

Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

355 360 365

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn

370 375 380

Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu

385 390 395 400

Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser

405 410 415

Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu

420 425 430

Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu

435 440 445

Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu

450 455 460

Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly

465 470 475 480

Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn

485 490 495

Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

500 505

<210> 57

<211> 534

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A141

<400> 57

Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg

1 5 10 15

Thr Asn Gln Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg

20 25 30

Leu His Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu

35 40 45

Val Lys Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln

50 55 60

Glu Cys Val Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys

65 70 75 80

Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly

85 90 95

Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly

100 105 110

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys

115 120 125

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu

130 135 140

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

145 150 155 160

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

165 170 175

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

180 185 190

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

195 200 205

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

210 215 220

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

225 230 235 240

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

245 250 255

Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln

260 265 270

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

275 280 285

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

290 295 300

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

305 310 315 320

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

325 330 335

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

340 345 350

Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

355 360 365

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

370 375 380

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

385 390 395 400

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

405 410 415

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

420 425 430

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

435 440 445

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

450 455 460

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

465 470 475 480

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

485 490 495

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

500 505 510

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

515 520 525

Asn Thr Ser Asn Pro Asp

530

<210> 58

<211> 714

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A142

<400> 58

Gly Asn Cys Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu

1 5 10 15

Tyr Lys Thr Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu

20 25 30

Ser Thr Ser Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys

35 40 45

Trp Met Ile Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu

50 55 60

Thr Cys Glu Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn

65 70 75 80

Lys Lys Asn Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile

85 90 95

Thr Trp Lys Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn

100 105 110

Glu Cys Ala Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu

115 120 125

Val Gln Tyr Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys

130 135 140

Pro Gly Ser Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys

145 150 155 160

Val Thr Cys Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr

165 170 175

Leu Cys Gly Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg

180 185 190

Lys Ala Thr Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly

195 200 205

Lys Cys Ile Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly

210 215 220

Lys Lys Cys Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu

225 230 235 240

Cys Asp Glu Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala

245 250 255

Ser Asp Asn Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala

260 265 270

Cys Ser Ser Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn

275 280 285

Ser Ile Ser Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp

290 295 300

Tyr Ser Phe Pro Ile Ser Ser Ile Leu Glu Trp Gly Gly Gly Gly Ser

305 310 315 320

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys Ser Cys Asp

325 330 335

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

340 345 350

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

355 360 365

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

370 375 380

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

385 390 395 400

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

405 410 415

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

420 425 430

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

435 440 445

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

450 455 460

Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu

465 470 475 480

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

485 490 495

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

500 505 510

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

515 520 525

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

530 535 540

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

545 550 555 560

Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

565 570 575

Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp Met Ile

580 585 590

Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe

595 600 605

Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser

610 615 620

Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val

625 630 635 640

Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys

645 650 655

His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala

660 665 670

Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe

675 680 685

Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe

690 695 700

Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

705 710

<210> 59

<211> 739

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A143

<400> 59

Gly Asn Cys Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu

1 5 10 15

Tyr Lys Thr Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu

20 25 30

Ser Thr Ser Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys

35 40 45

Trp Met Ile Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu

50 55 60

Thr Cys Glu Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn

65 70 75 80

Lys Lys Asn Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile

85 90 95

Thr Trp Lys Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn

100 105 110

Glu Cys Ala Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu

115 120 125

Val Gln Tyr Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys

130 135 140

Pro Gly Ser Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys

145 150 155 160

Val Thr Cys Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr

165 170 175

Leu Cys Gly Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg

180 185 190

Lys Ala Thr Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly

195 200 205

Lys Cys Ile Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly

210 215 220

Lys Lys Cys Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu

225 230 235 240

Cys Asp Glu Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala

245 250 255

Ser Asp Asn Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala

260 265 270

Cys Ser Ser Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn

275 280 285

Ser Ile Ser Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp

290 295 300

Tyr Ser Phe Pro Ile Ser Ser Ile Leu Glu Trp Gly Gly Gly Gly Ser

305 310 315 320

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys Ser Cys Asp

325 330 335

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

340 345 350

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

355 360 365

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

370 375 380

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

385 390 395 400

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

405 410 415

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

420 425 430

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

435 440 445

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

450 455 460

Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu

465 470 475 480

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

485 490 495

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

500 505 510

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

515 520 525

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

530 535 540

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

545 550 555 560

Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

565 570 575

Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met Glu Ala

580 585 590

Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala His Pro

595 600 605

Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala

610 615 620

Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr

625 630 635 640

Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile

645 650 655

Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp

660 665 670

Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr

675 680 685

His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys

690 695 700

Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser

705 710 715 720

Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser

725 730 735

Asn Pro Asp

<210> 60

<211> 714

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A117

<400> 60

Gly Asn Cys Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu

1 5 10 15

Tyr Lys Thr Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu

20 25 30

Ser Thr Ser Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys

35 40 45

Trp Met Ile Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu

50 55 60

Thr Cys Glu Asn Val Asp Cys Gly Pro Gly Gln Ser Cys Val Val Asp

65 70 75 80

Gln Thr Gly Ser Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile

85 90 95

Thr Trp Lys Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn

100 105 110

Glu Cys Ala Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu

115 120 125

Val Gln Tyr Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys

130 135 140

Pro Gly Ser Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys

145 150 155 160

Val Thr Cys Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr

165 170 175

Leu Cys Gly Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg

180 185 190

Lys Ala Thr Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly

195 200 205

Lys Cys Ile Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly

210 215 220

Lys Lys Cys Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu

225 230 235 240

Cys Asp Glu Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala

245 250 255

Ser Asp Asn Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala

260 265 270

Cys Ser Ser Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn

275 280 285

Ser Ile Ser Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp

290 295 300

Tyr Ser Phe Pro Ile Ser Ser Ile Leu Glu Trp Gly Gly Gly Gly Ser

305 310 315 320

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys Ser Cys Asp

325 330 335

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

340 345 350

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

355 360 365

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

370 375 380

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

385 390 395 400

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

405 410 415

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

420 425 430

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

435 440 445

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

450 455 460

Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu

465 470 475 480

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

485 490 495

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

500 505 510

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

515 520 525

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

530 535 540

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

545 550 555 560

Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

565 570 575

Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp Met Ile

580 585 590

Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe

595 600 605

Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser

610 615 620

Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val

625 630 635 640

Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys

645 650 655

His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala

660 665 670

Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe

675 680 685

Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe

690 695 700

Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

705 710

<210> 61

<211> 739

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A118

<400> 61

Gly Asn Cys Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu

1 5 10 15

Tyr Lys Thr Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu

20 25 30

Ser Thr Ser Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys

35 40 45

Trp Met Ile Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu

50 55 60

Thr Cys Glu Asn Val Asp Cys Gly Pro Gly Gln Ser Cys Val Val Asp

65 70 75 80

Gln Thr Gly Ser Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile

85 90 95

Thr Trp Lys Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn

100 105 110

Glu Cys Ala Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu

115 120 125

Val Gln Tyr Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys

130 135 140

Pro Gly Ser Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys

145 150 155 160

Val Thr Cys Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr

165 170 175

Leu Cys Gly Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg

180 185 190

Lys Ala Thr Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly

195 200 205

Lys Cys Ile Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly

210 215 220

Lys Lys Cys Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu

225 230 235 240

Cys Asp Glu Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala

245 250 255

Ser Asp Asn Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala

260 265 270

Cys Ser Ser Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn

275 280 285

Ser Ile Ser Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp

290 295 300

Tyr Ser Phe Pro Ile Ser Ser Ile Leu Glu Trp Gly Gly Gly Gly Ser

305 310 315 320

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys Ser Cys Asp

325 330 335

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

340 345 350

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

355 360 365

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

370 375 380

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

385 390 395 400

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

405 410 415

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

420 425 430

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

435 440 445

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

450 455 460

Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu

465 470 475 480

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

485 490 495

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

500 505 510

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

515 520 525

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

530 535 540

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

545 550 555 560

Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

565 570 575

Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met Glu Ala

580 585 590

Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala His Pro

595 600 605

Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala

610 615 620

Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr

625 630 635 640

Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile

645 650 655

Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp

660 665 670

Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr

675 680 685

His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys

690 695 700

Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser

705 710 715 720

Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser

725 730 735

Asn Pro Asp

<210> 62

<211> 687

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A144

<400> 62

Gly Asn Cys Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu

1 5 10 15

Tyr Lys Thr Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu

20 25 30

Ser Thr Ser Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys

35 40 45

Trp Met Ile Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu

50 55 60

Thr Cys Glu Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn

65 70 75 80

Lys Lys Asn Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile

85 90 95

Thr Trp Lys Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn

100 105 110

Glu Cys Ala Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu

115 120 125

Val Gln Tyr Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys

130 135 140

Pro Gly Ser Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys

145 150 155 160

Val Thr Cys Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr

165 170 175

Leu Cys Gly Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg

180 185 190

Lys Ala Thr Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly

195 200 205

Lys Cys Ile Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly

210 215 220

Lys Lys Cys Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu

225 230 235 240

Cys Asp Glu Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala

245 250 255

Ser Asp Asn Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala

260 265 270

Cys Ser Ser Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn

275 280 285

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu

290 295 300

Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro

305 310 315 320

Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

325 330 335

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

340 345 350

Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp

355 360 365

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr

370 375 380

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

385 390 395 400

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu

405 410 415

Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

420 425 430

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys

435 440 445

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

450 455 460

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

465 470 475 480

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

485 490 495

Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser

500 505 510

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

515 520 525

Leu Ser Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly

530 535 540

Ser Gly Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val

545 550 555 560

Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro

565 570 575

Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln

580 585 590

Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro

595 600 605

Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr

610 615 620

Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile

625 630 635 640

Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys

645 650 655

Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn

660 665 670

Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

675 680 685

<210> 63

<211> 712

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of A145

<400> 63

Gly Asn Cys Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu

1 5 10 15

Tyr Lys Thr Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu

20 25 30

Ser Thr Ser Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys

35 40 45

Trp Met Ile Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu

50 55 60

Thr Cys Glu Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn

65 70 75 80

Lys Lys Asn Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile

85 90 95

Thr Trp Lys Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn

100 105 110

Glu Cys Ala Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu

115 120 125

Val Gln Tyr Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys

130 135 140

Pro Gly Ser Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys

145 150 155 160

Val Thr Cys Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr

165 170 175

Leu Cys Gly Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg

180 185 190

Lys Ala Thr Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly

195 200 205

Lys Cys Ile Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly

210 215 220

Lys Lys Cys Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu

225 230 235 240

Cys Asp Glu Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala

245 250 255

Ser Asp Asn Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala

260 265 270

Cys Ser Ser Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn

275 280 285

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu

290 295 300

Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro

305 310 315 320

Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

325 330 335

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

340 345 350

Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp

355 360 365

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr

370 375 380

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

385 390 395 400

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu

405 410 415

Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

420 425 430

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys

435 440 445

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

450 455 460

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

465 470 475 480

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

485 490 495

Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser

500 505 510

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

515 520 525

Leu Ser Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly

530 535 540

Ser Gly Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp

545 550 555 560

Val Glu Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn

565 570 575

Arg Thr Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr

580 585 590

Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp

595 600 605

Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys

610 615 620

Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val

625 630 635 640

Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp

645 650 655

Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro

660 665 670

Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met

675 680 685

Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu

690 695 700

Glu Tyr Asn Thr Ser Asn Pro Asp

705 710

<210> 64

<211> 509

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A146

<400> 64

Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg

1 5 10 15

Thr Asn Gln Ser Gly Leu Glu Arg Cys Glu Gly Asp Gln Asp Lys Arg

20 25 30

Leu His Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu

35 40 45

Val Lys Lys Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Gln

50 55 60

Glu Cys Val Ala Thr Lys Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys

65 70 75 80

Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly

85 90 95

Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly

100 105 110

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys

115 120 125

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu

130 135 140

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

145 150 155 160

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

165 170 175

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

180 185 190

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

195 200 205

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

210 215 220

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

225 230 235 240

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

245 250 255

Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln

260 265 270

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

275 280 285

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

290 295 300

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

305 310 315 320

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

325 330 335

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

340 345 350

Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

355 360 365

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn

370 375 380

Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu

385 390 395 400

Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser

405 410 415

Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu

420 425 430

Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu

435 440 445

Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu

450 455 460

Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly

465 470 475 480

Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn

485 490 495

Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

500 505

<210> 65

<211> 534

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A147

<400> 65

Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg

1 5 10 15

Thr Asn Gln Ser Gly Leu Glu Arg Cys Glu Gly Asp Gln Asp Lys Arg

20 25 30

Leu His Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu

35 40 45

Val Lys Lys Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Gln

50 55 60

Glu Cys Val Ala Thr Lys Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys

65 70 75 80

Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly

85 90 95

Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly

100 105 110

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys

115 120 125

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu

130 135 140

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

145 150 155 160

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

165 170 175

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

180 185 190

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

195 200 205

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

210 215 220

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

225 230 235 240

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

245 250 255

Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln

260 265 270

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

275 280 285

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

290 295 300

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

305 310 315 320

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

325 330 335

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

340 345 350

Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

355 360 365

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

370 375 380

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

385 390 395 400

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

405 410 415

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

420 425 430

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

435 440 445

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

450 455 460

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

465 470 475 480

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

485 490 495

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

500 505 510

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

515 520 525

Asn Thr Ser Asn Pro Asp

530

<210> 66

<211> 509

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A148

<400> 66

Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg

1 5 10 15

Thr Asn Gln Ser Gly Leu Glu Arg Cys Tyr Gly Asp Lys Asp Lys Arg

20 25 30

Arg His Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu

35 40 45

Val Lys Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln

50 55 60

Glu Cys Val Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys

65 70 75 80

Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly

85 90 95

Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly

100 105 110

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys

115 120 125

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu

130 135 140

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

145 150 155 160

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

165 170 175

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

180 185 190

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

195 200 205

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

210 215 220

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

225 230 235 240

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

245 250 255

Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln

260 265 270

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

275 280 285

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

290 295 300

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

305 310 315 320

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

325 330 335

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

340 345 350

Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

355 360 365

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn

370 375 380

Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu

385 390 395 400

Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser

405 410 415

Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu

420 425 430

Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu

435 440 445

Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu

450 455 460

Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly

465 470 475 480

Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn

485 490 495

Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

500 505

<210> 67

<211> 534

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A149

<400> 67

Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg

1 5 10 15

Thr Asn Gln Ser Gly Leu Glu Arg Cys Tyr Gly Asp Lys Asp Lys Arg

20 25 30

Arg His Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu

35 40 45

Val Lys Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln

50 55 60

Glu Cys Val Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys

65 70 75 80

Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly

85 90 95

Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly

100 105 110

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys

115 120 125

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu

130 135 140

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

145 150 155 160

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

165 170 175

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

180 185 190

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

195 200 205

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

210 215 220

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

225 230 235 240

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

245 250 255

Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln

260 265 270

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

275 280 285

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

290 295 300

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

305 310 315 320

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

325 330 335

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

340 345 350

Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

355 360 365

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

370 375 380

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

385 390 395 400

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

405 410 415

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

420 425 430

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

435 440 445

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

450 455 460

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

465 470 475 480

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

485 490 495

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

500 505 510

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

515 520 525

Asn Thr Ser Asn Pro Asp

530

<210> 68

<211> 506

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A150

<400> 68

Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg

1 5 10 15

Thr Asn Gln Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg

20 25 30

Leu His Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu

35 40 45

Val Lys Lys Gly Cys Trp Asp Asp Asp Phe Asn Cys Tyr Asp Arg Gln

50 55 60

Glu Cys Val Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys

65 70 75 80

Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly

85 90 95

Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro Thr Gly Gly Gly Gly Ser

100 105 110

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys Ser Cys Asp

115 120 125

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

130 135 140

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

145 150 155 160

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

165 170 175

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

180 185 190

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

195 200 205

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

210 215 220

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

225 230 235 240

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

245 250 255

Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu

260 265 270

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

275 280 285

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

290 295 300

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

305 310 315 320

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

325 330 335

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

340 345 350

Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

355 360 365

Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp Met Ile

370 375 380

Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe

385 390 395 400

Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser

405 410 415

Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val

420 425 430

Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys

435 440 445

His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala

450 455 460

Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe

465 470 475 480

Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe

485 490 495

Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

500 505

<210> 69

<211> 531

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of A151

<400> 69

Glu Thr Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg

1 5 10 15

Thr Asn Gln Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg

20 25 30

Leu His Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu

35 40 45

Val Lys Lys Gly Cys Trp Asp Asp Asp Phe Asn Cys Tyr Asp Arg Gln

50 55 60

Glu Cys Val Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys

65 70 75 80

Glu Gly Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly

85 90 95

Gly Pro Glu Val Thr Tyr Glu Pro Pro Pro Thr Gly Gly Gly Gly Ser

100 105 110

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys Ser Cys Asp

115 120 125

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

130 135 140

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

145 150 155 160

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

165 170 175

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

180 185 190

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

195 200 205

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

210 215 220

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

225 230 235 240

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

245 250 255

Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu

260 265 270

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

275 280 285

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

290 295 300

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

305 310 315 320

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

325 330 335

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

340 345 350

Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

355 360 365

Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met Glu Ala

370 375 380

Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala His Pro

385 390 395 400

Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala

405 410 415

Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr

420 425 430

Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile

435 440 445

Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp

450 455 460

Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr

465 470 475 480

His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys

485 490 495

Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser

500 505 510

Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser

515 520 525

Asn Pro Asp

530

<210> 70

<211> 508

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of A152

<400> 70

Gly Ala Ile Leu Gly Arg Ser Glu Thr Gln Glu Cys Leu Phe Tyr Asn

1 5 10 15

Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln Thr Gly Val Glu Pro Cys

20 25 30

Glu Gly Glu Lys Asp Lys Arg Leu His Cys Tyr Ala Thr Trp Arg Asn

35 40 45

Ile Ser Gly Ser Ile Glu Ile Val Lys Lys Gly Cys Trp Leu Asp Asp

50 55 60

Phe Asn Cys Tyr Asp Arg Thr Asp Cys Val Glu Thr Glu Glu Asn Pro

65 70 75 80

Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn Met Cys Asn Glu Lys Phe

85 90 95

Ser Tyr Phe Pro Glu Met Glu Val Thr Gln Pro Thr Ser Gly Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys Ser

115 120 125

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

130 135 140

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

145 150 155 160

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

165 170 175

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

180 185 190

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

195 200 205

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

210 215 220

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

225 230 235 240

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

245 250 255

Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val

260 265 270

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

275 280 285

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

290 295 300

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

305 310 315 320

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

325 330 335

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

340 345 350

Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

355 360 365

Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp

370 375 380

Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys

385 390 395 400

Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys

405 410 415

Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val

420 425 430

Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr

435 440 445

Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp

450 455 460

Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu

465 470 475 480

Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile

485 490 495

Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

500 505

<210> 71

<211> 533

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence of A153

<400> 71

Gly Ala Ile Leu Gly Arg Ser Glu Thr Gln Glu Cys Leu Phe Tyr Asn

1 5 10 15

Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln Thr Gly Val Glu Pro Cys

20 25 30

Glu Gly Glu Lys Asp Lys Arg Leu His Cys Tyr Ala Thr Trp Arg Asn

35 40 45

Ile Ser Gly Ser Ile Glu Ile Val Lys Lys Gly Cys Trp Leu Asp Asp

50 55 60

Phe Asn Cys Tyr Asp Arg Thr Asp Cys Val Glu Thr Glu Glu Asn Pro

65 70 75 80

Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn Met Cys Asn Glu Lys Phe

85 90 95

Ser Tyr Phe Pro Glu Met Glu Val Thr Gln Pro Thr Ser Gly Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys Ser

115 120 125

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

130 135 140

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

145 150 155 160

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

165 170 175

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

180 185 190

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

195 200 205

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

210 215 220

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

225 230 235 240

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

245 250 255

Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val

260 265 270

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

275 280 285

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

290 295 300

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

305 310 315 320

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

325 330 335

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

340 345 350

Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

355 360 365

Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met

370 375 380

Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala

385 390 395 400

His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn

405 410 415

Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe

420 425 430

Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr

435 440 445

Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys

450 455 460

Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu

465 470 475 480

Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile

485 490 495

Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys

500 505 510

Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn

515 520 525

Thr Ser Asn Pro Asp

530

<210> 72

<211> 604

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A154

<400> 72

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Gly Leu Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ile Pro Tyr Asn Gly Asn Thr Asn Ser Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Arg Asp Arg Asp Tyr Gly Val Asn Tyr Asp Ala Phe Asp Ile Trp

100 105 110

Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

225 230 235 240

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Val Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

450 455 460

Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp

465 470 475 480

Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys

485 490 495

Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys

500 505 510

Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val

515 520 525

Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr

530 535 540

Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp

545 550 555 560

Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu

565 570 575

Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile

580 585 590

Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

595 600

<210> 73

<211> 629

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A155

<400> 73

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Gly Leu Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ile Pro Tyr Asn Gly Asn Thr Asn Ser Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Arg Asp Arg Asp Tyr Gly Val Asn Tyr Asp Ala Phe Asp Ile Trp

100 105 110

Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

225 230 235 240

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Val Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

450 455 460

Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met

465 470 475 480

Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala

485 490 495

His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn

500 505 510

Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe

515 520 525

Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr

530 535 540

Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys

545 550 555 560

Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu

565 570 575

Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile

580 585 590

Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys

595 600 605

Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn

610 615 620

Thr Ser Asn Pro Asp

625

<210> 74

<211> 605

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A156

<400> 74

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Phe Ser Ser His

20 25 30

Phe Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Leu Tyr Thr Gly Gly Thr Ser Phe Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Ser Met Ser Val Gly Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Ala Arg Ser Gly Ile Thr Phe Thr Gly Ile Ile Val Pro Gly Ser

100 105 110

Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser

115 120 125

Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr

130 135 140

Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro

145 150 155 160

Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val

165 170 175

His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser

180 185 190

Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr

195 200 205

Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val

210 215 220

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe

225 230 235 240

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

245 250 255

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

260 265 270

Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val

275 280 285

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser

290 295 300

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

305 310 315 320

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser

325 330 335

Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

340 345 350

Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu

405 410 415

Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Leu Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

450 455 460

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn

465 470 475 480

Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu

485 490 495

Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser

500 505 510

Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu

515 520 525

Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu

530 535 540

Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu

545 550 555 560

Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly

565 570 575

Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn

580 585 590

Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

595 600 605

<210> 75

<211> 630

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A157

<400> 75

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Phe Ser Ser His

20 25 30

Phe Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Leu Tyr Thr Gly Gly Thr Ser Phe Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Ser Met Ser Val Gly Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Ala Arg Ser Gly Ile Thr Phe Thr Gly Ile Ile Val Pro Gly Ser

100 105 110

Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser

115 120 125

Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr

130 135 140

Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro

145 150 155 160

Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val

165 170 175

His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser

180 185 190

Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr

195 200 205

Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val

210 215 220

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe

225 230 235 240

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

245 250 255

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

260 265 270

Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val

275 280 285

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser

290 295 300

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

305 310 315 320

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser

325 330 335

Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

340 345 350

Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu

405 410 415

Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Leu Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

450 455 460

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

465 470 475 480

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

485 490 495

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

500 505 510

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

515 520 525

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

530 535 540

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

545 550 555 560

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

565 570 575

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

580 585 590

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

595 600 605

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

610 615 620

Asn Thr Ser Asn Pro Asp

625 630

<210> 76

<211> 572

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A158

<400> 76

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Thr

450 455 460

Gln Glu Cys Leu Phe Phe Asn Ala Asn Trp Glu Lys Asp Arg Thr Asn

465 470 475 480

Gln Thr Gly Val Glu Pro Cys Tyr Gly Asp Lys Asp Lys Arg Arg His

485 490 495

Cys Phe Ala Thr Trp Lys Asn Ile Ser Gly Ser Ile Glu Ile Val Lys

500 505 510

Gln Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Thr Asp Cys

515 520 525

Val Glu Lys Lys Asp Ser Pro Glu Val Tyr Phe Cys Cys Cys Glu Gly

530 535 540

Asn Met Cys Asn Glu Lys Phe Ser Tyr Phe Pro Glu Met Glu Val Thr

545 550 555 560

Gln Pro Thr Ser Asn Pro Val Thr Pro Lys Pro Pro

565 570

<210> 77

<211> 572

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A159

<400> 77

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Thr

450 455 460

Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn

465 470 475 480

Gln Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg Leu His

485 490 495

Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys

500 505 510

Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys

515 520 525

Val Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly

530 535 540

Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro

545 550 555 560

Glu Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr

565 570

<210> 78

<211> 777

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A160

<400> 78

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Asn

450 455 460

Cys Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys

465 470 475 480

Thr Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr

485 490 495

Ser Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met

500 505 510

Ile Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys

515 520 525

Glu Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn Lys Lys

530 535 540

Asn Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp

545 550 555 560

Lys Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys

565 570 575

Ala Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln

580 585 590

Tyr Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly

595 600 605

Ser Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr

610 615 620

Cys Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys

625 630 635 640

Gly Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala

645 650 655

Thr Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys

660 665 670

Ile Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys

675 680 685

Cys Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp

690 695 700

Glu Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp

705 710 715 720

Asn Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser

725 730 735

Ser Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Ser Ile

740 745 750

Ser Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp Tyr Ser

755 760 765

Phe Pro Ile Ser Ser Ile Leu Glu Trp

770 775

<210> 79

<211> 777

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A161

<400> 79

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Asn

450 455 460

Cys Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys

465 470 475 480

Thr Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr

485 490 495

Ser Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met

500 505 510

Ile Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys

515 520 525

Glu Asn Val Asp Cys Gly Pro Gly Gln Ser Cys Val Val Asp Gln Thr

530 535 540

Gly Ser Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp

545 550 555 560

Lys Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys

565 570 575

Ala Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln

580 585 590

Tyr Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly

595 600 605

Ser Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr

610 615 620

Cys Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys

625 630 635 640

Gly Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala

645 650 655

Thr Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys

660 665 670

Ile Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys

675 680 685

Cys Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp

690 695 700

Glu Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp

705 710 715 720

Asn Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser

725 730 735

Ser Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Ser Ile

740 745 750

Ser Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp Tyr Ser

755 760 765

Phe Pro Ile Ser Ser Ile Leu Glu Trp

770 775

<210> 80

<211> 777

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A162

<400> 80

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Asn

450 455 460

Cys Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys

465 470 475 480

Thr Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr

485 490 495

Ser Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met

500 505 510

Ile Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys

515 520 525

Glu Asn Val Asp Cys Gly Pro Gly Gln Ser Cys Val Val Asp Gln Thr

530 535 540

Gly Ser Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp

545 550 555 560

Lys Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys

565 570 575

Ala Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln

580 585 590

Tyr Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly

595 600 605

Ser Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr

610 615 620

Cys Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys

625 630 635 640

Gly Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala

645 650 655

Thr Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys

660 665 670

Ile Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys

675 680 685

Cys Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp

690 695 700

Glu Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp

705 710 715 720

Asn Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser

725 730 735

Ser Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Ser Ile

740 745 750

Ser Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp Tyr Ser

755 760 765

Phe Pro Ile Ser Ser Ile Leu Glu Trp

770 775

<210> 81

<211> 572

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A163

<400> 81

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Thr

450 455 460

Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn

465 470 475 480

Gln Ser Gly Leu Glu Arg Cys Glu Gly Asp Gln Asp Lys Arg Leu His

485 490 495

Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys

500 505 510

Lys Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Gln Glu Cys

515 520 525

Val Ala Thr Lys Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly

530 535 540

Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro

545 550 555 560

Glu Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr

565 570

<210> 82

<211> 572

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A164

<400> 82

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Thr

450 455 460

Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn

465 470 475 480

Gln Ser Gly Leu Glu Arg Cys Tyr Gly Asp Lys Asp Lys Arg Arg His

485 490 495

Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys

500 505 510

Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys

515 520 525

Val Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly

530 535 540

Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro

545 550 555 560

Glu Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr

565 570

<210> 83

<211> 569

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A165

<400> 83

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Thr

450 455 460

Arg Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn

465 470 475 480

Gln Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg Leu His

485 490 495

Cys Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys

500 505 510

Lys Gly Cys Trp Asp Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys

515 520 525

Val Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly

530 535 540

Asn Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro

545 550 555 560

Glu Val Thr Tyr Glu Pro Pro Pro Thr

565

<210> 84

<211> 571

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A166

<400> 84

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Ala

450 455 460

Ile Leu Gly Arg Ser Glu Thr Gln Glu Cys Leu Phe Tyr Asn Ala Asn

465 470 475 480

Trp Glu Leu Glu Arg Thr Asn Gln Thr Gly Val Glu Pro Cys Glu Gly

485 490 495

Glu Lys Asp Lys Arg Leu His Cys Tyr Ala Thr Trp Arg Asn Ile Ser

500 505 510

Gly Ser Ile Glu Ile Val Lys Lys Gly Cys Trp Leu Asp Asp Phe Asn

515 520 525

Cys Tyr Asp Arg Thr Asp Cys Val Glu Thr Glu Glu Asn Pro Gln Val

530 535 540

Tyr Phe Cys Cys Cys Glu Gly Asn Met Cys Asn Glu Lys Phe Ser Tyr

545 550 555 560

Phe Pro Glu Met Glu Val Thr Gln Pro Thr Ser

565 570

<210> 85

<211> 733

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A169

<400> 85

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Gln

450 455 460

Glu Cys Leu Phe Phe Asn Ala Asn Trp Glu Lys Asp Arg Thr Asn Gln

465 470 475 480

Thr Gly Val Glu Pro Cys Tyr Gly Asp Lys Asp Lys Arg Arg His Cys

485 490 495

Phe Ala Thr Trp Lys Asn Ile Ser Gly Ser Ile Glu Ile Val Lys Gln

500 505 510

Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Thr Asp Cys Val

515 520 525

Glu Lys Lys Asp Ser Pro Glu Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Met Cys Asn Glu Lys Phe Ser Tyr Phe Pro Glu Met Glu Val Thr Gln

545 550 555 560

Pro Thr Ser Asn Pro Val Thr Pro Lys Pro Pro Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn

595 600 605

Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu

610 615 620

Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser

625 630 635 640

Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu

645 650 655

Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu

660 665 670

Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu

675 680 685

Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly

690 695 700

Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn

705 710 715 720

Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

725 730

<210> 86

<211> 758

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A170

<400> 86

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Gln

450 455 460

Glu Cys Leu Phe Phe Asn Ala Asn Trp Glu Lys Asp Arg Thr Asn Gln

465 470 475 480

Thr Gly Val Glu Pro Cys Tyr Gly Asp Lys Asp Lys Arg Arg His Cys

485 490 495

Phe Ala Thr Trp Lys Asn Ile Ser Gly Ser Ile Glu Ile Val Lys Gln

500 505 510

Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Thr Asp Cys Val

515 520 525

Glu Lys Lys Asp Ser Pro Glu Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Met Cys Asn Glu Lys Phe Ser Tyr Phe Pro Glu Met Glu Val Thr Gln

545 550 555 560

Pro Thr Ser Asn Pro Val Thr Pro Lys Pro Pro Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

595 600 605

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

610 615 620

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

625 630 635 640

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

645 650 655

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

660 665 670

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

675 680 685

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

690 695 700

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

705 710 715 720

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

725 730 735

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

740 745 750

Asn Thr Ser Asn Pro Asp

755

<210> 87

<211> 733

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A171

<400> 87

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg Leu His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn

595 600 605

Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu

610 615 620

Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser

625 630 635 640

Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu

645 650 655

Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu

660 665 670

Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu

675 680 685

Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly

690 695 700

Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn

705 710 715 720

Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

725 730

<210> 88

<211> 758

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A172

<400> 88

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg Leu His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

595 600 605

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

610 615 620

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

625 630 635 640

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

645 650 655

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

660 665 670

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

675 680 685

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

690 695 700

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

705 710 715 720

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

725 730 735

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

740 745 750

Asn Thr Ser Asn Pro Asp

755

<210> 89

<211> 938

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A173

<400> 89

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Asn Cys

450 455 460

Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys Thr

465 470 475 480

Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr Ser

485 490 495

Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met Ile

500 505 510

Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys Glu

515 520 525

Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn Lys Lys Asn

530 535 540

Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp Lys

545 550 555 560

Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys Ala

565 570 575

Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln Tyr

580 585 590

Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly Ser

595 600 605

Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr Cys

610 615 620

Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys Gly

625 630 635 640

Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala Thr

645 650 655

Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys Ile

660 665 670

Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys Cys

675 680 685

Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp Glu

690 695 700

Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp Asn

705 710 715 720

Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser Ser

725 730 735

Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Ser Ile Ser

740 745 750

Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp Tyr Ser Phe

755 760 765

Pro Ile Ser Ser Ile Leu Glu Trp Gly Gly Gly Gly Ser Gly Gly Gly

770 775 780

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

785 790 795 800

Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp Met Ile

805 810 815

Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe

820 825 830

Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser

835 840 845

Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val

850 855 860

Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys

865 870 875 880

His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala

885 890 895

Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe

900 905 910

Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe

915 920 925

Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

930 935

<210> 90

<211> 963

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A174

<400> 90

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Asn Cys

450 455 460

Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys Thr

465 470 475 480

Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr Ser

485 490 495

Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met Ile

500 505 510

Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys Glu

515 520 525

Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn Lys Lys Asn

530 535 540

Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp Lys

545 550 555 560

Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys Ala

565 570 575

Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln Tyr

580 585 590

Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly Ser

595 600 605

Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr Cys

610 615 620

Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys Gly

625 630 635 640

Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala Thr

645 650 655

Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys Ile

660 665 670

Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys Cys

675 680 685

Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp Glu

690 695 700

Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp Asn

705 710 715 720

Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser Ser

725 730 735

Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Ser Ile Ser

740 745 750

Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp Tyr Ser Phe

755 760 765

Pro Ile Ser Ser Ile Leu Glu Trp Gly Gly Gly Gly Ser Gly Gly Gly

770 775 780

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

785 790 795 800

Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met Glu Ala

805 810 815

Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala His Pro

820 825 830

Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala

835 840 845

Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr

850 855 860

Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile

865 870 875 880

Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp

885 890 895

Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr

900 905 910

His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys

915 920 925

Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser

930 935 940

Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser

945 950 955 960

Asn Pro Asp

<210> 91

<211> 938

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A175

<400> 91

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Asn Cys

450 455 460

Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys Thr

465 470 475 480

Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr Ser

485 490 495

Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met Ile

500 505 510

Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys Glu

515 520 525

Asn Val Asp Cys Gly Pro Gly Gln Ser Cys Val Val Asp Gln Thr Gly

530 535 540

Ser Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp Lys

545 550 555 560

Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys Ala

565 570 575

Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln Tyr

580 585 590

Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly Ser

595 600 605

Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr Cys

610 615 620

Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys Gly

625 630 635 640

Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala Thr

645 650 655

Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys Ile

660 665 670

Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys Cys

675 680 685

Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp Glu

690 695 700

Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp Asn

705 710 715 720

Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser Ser

725 730 735

Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Ser Ile Ser

740 745 750

Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp Tyr Ser Phe

755 760 765

Pro Ile Ser Ser Ile Leu Glu Trp Gly Gly Gly Gly Ser Gly Gly Gly

770 775 780

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

785 790 795 800

Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp Met Ile

805 810 815

Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe

820 825 830

Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser

835 840 845

Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val

850 855 860

Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys

865 870 875 880

His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala

885 890 895

Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe

900 905 910

Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe

915 920 925

Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

930 935

<210> 92

<211> 963

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A176

<400> 92

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Asn Cys

450 455 460

Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys Thr

465 470 475 480

Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr Ser

485 490 495

Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met Ile

500 505 510

Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys Glu

515 520 525

Asn Val Asp Cys Gly Pro Gly Gln Ser Cys Val Val Asp Gln Thr Gly

530 535 540

Ser Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp Lys

545 550 555 560

Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys Ala

565 570 575

Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln Tyr

580 585 590

Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly Ser

595 600 605

Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr Cys

610 615 620

Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys Gly

625 630 635 640

Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala Thr

645 650 655

Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys Ile

660 665 670

Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys Cys

675 680 685

Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp Glu

690 695 700

Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp Asn

705 710 715 720

Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser Ser

725 730 735

Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Ser Ile Ser

740 745 750

Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp Tyr Ser Phe

755 760 765

Pro Ile Ser Ser Ile Leu Glu Trp Gly Gly Gly Gly Ser Gly Gly Gly

770 775 780

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

785 790 795 800

Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met Glu Ala

805 810 815

Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala His Pro

820 825 830

Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala

835 840 845

Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr

850 855 860

Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile

865 870 875 880

Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp

885 890 895

Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr

900 905 910

His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys

915 920 925

Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser

930 935 940

Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser

945 950 955 960

Asn Pro Asp

<210> 93

<211> 911

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A177

<400> 93

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Asn Cys

450 455 460

Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys Thr

465 470 475 480

Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr Ser

485 490 495

Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met Ile

500 505 510

Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys Glu

515 520 525

Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn Lys Lys Asn

530 535 540

Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp Lys

545 550 555 560

Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys Ala

565 570 575

Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln Tyr

580 585 590

Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly Ser

595 600 605

Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr Cys

610 615 620

Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys Gly

625 630 635 640

Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala Thr

645 650 655

Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys Ile

660 665 670

Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys Cys

675 680 685

Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp Glu

690 695 700

Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp Asn

705 710 715 720

Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser Ser

725 730 735

Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Gly Gly Gly

740 745 750

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

755 760 765

Ser Gly Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val

770 775 780

Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro

785 790 795 800

Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln

805 810 815

Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro

820 825 830

Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr

835 840 845

Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile

850 855 860

Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys

865 870 875 880

Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn

885 890 895

Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

900 905 910

<210> 94

<211> 936

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A178

<400> 94

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Asn Cys

450 455 460

Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys Thr

465 470 475 480

Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr Ser

485 490 495

Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met Ile

500 505 510

Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys Glu

515 520 525

Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn Lys Lys Asn

530 535 540

Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp Lys

545 550 555 560

Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys Ala

565 570 575

Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln Tyr

580 585 590

Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly Ser

595 600 605

Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr Cys

610 615 620

Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys Gly

625 630 635 640

Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala Thr

645 650 655

Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys Ile

660 665 670

Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys Cys

675 680 685

Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp Glu

690 695 700

Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp Asn

705 710 715 720

Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser Ser

725 730 735

Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Gly Gly Gly

740 745 750

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

755 760 765

Ser Gly Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp

770 775 780

Val Glu Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn

785 790 795 800

Arg Thr Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr

805 810 815

Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp

820 825 830

Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys

835 840 845

Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val

850 855 860

Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp

865 870 875 880

Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro

885 890 895

Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met

900 905 910

Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu

915 920 925

Glu Tyr Asn Thr Ser Asn Pro Asp

930 935

<210> 95

<211> 733

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A179

<400> 95

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Glu Gly Asp Gln Asp Lys Arg Leu His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Lys Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn

595 600 605

Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu

610 615 620

Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser

625 630 635 640

Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu

645 650 655

Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu

660 665 670

Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu

675 680 685

Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly

690 695 700

Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn

705 710 715 720

Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

725 730

<210> 96

<211> 758

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A180

<400> 96

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Glu Gly Asp Gln Asp Lys Arg Leu His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Lys Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

595 600 605

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

610 615 620

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

625 630 635 640

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

645 650 655

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

660 665 670

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

675 680 685

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

690 695 700

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

705 710 715 720

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

725 730 735

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

740 745 750

Asn Thr Ser Asn Pro Asp

755

<210> 97

<211> 733

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A181

<400> 97

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Tyr Gly Asp Lys Asp Lys Arg Arg His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn

595 600 605

Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu

610 615 620

Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser

625 630 635 640

Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu

645 650 655

Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu

660 665 670

Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu

675 680 685

Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly

690 695 700

Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn

705 710 715 720

Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

725 730

<210> 98

<211> 758

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A182

<400> 98

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Tyr Gly Asp Lys Asp Lys Arg Arg His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

595 600 605

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

610 615 620

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

625 630 635 640

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

645 650 655

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

660 665 670

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

675 680 685

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

690 695 700

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

705 710 715 720

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

725 730 735

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

740 745 750

Asn Thr Ser Asn Pro Asp

755

<210> 99

<211> 730

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A183

<400> 99

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg Leu His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Asp Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Gly Gly Gly Gly Ser Gly Gly Gly

565 570 575

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

580 585 590

Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp Met Ile

595 600 605

Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe

610 615 620

Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser

625 630 635 640

Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val

645 650 655

Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys

660 665 670

His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala

675 680 685

Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe

690 695 700

Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe

705 710 715 720

Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

725 730

<210> 100

<211> 755

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A184

<400> 100

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg Leu His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Asp Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Gly Gly Gly Gly Ser Gly Gly Gly

565 570 575

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

580 585 590

Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met Glu Ala

595 600 605

Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala His Pro

610 615 620

Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala

625 630 635 640

Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr

645 650 655

Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile

660 665 670

Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp

675 680 685

Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr

690 695 700

His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys

705 710 715 720

Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser

725 730 735

Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser

740 745 750

Asn Pro Asp

755

<210> 101

<211> 732

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A185

<400> 101

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ala Ile

450 455 460

Leu Gly Arg Ser Glu Thr Gln Glu Cys Leu Phe Tyr Asn Ala Asn Trp

465 470 475 480

Glu Leu Glu Arg Thr Asn Gln Thr Gly Val Glu Pro Cys Glu Gly Glu

485 490 495

Lys Asp Lys Arg Leu His Cys Tyr Ala Thr Trp Arg Asn Ile Ser Gly

500 505 510

Ser Ile Glu Ile Val Lys Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys

515 520 525

Tyr Asp Arg Thr Asp Cys Val Glu Thr Glu Glu Asn Pro Gln Val Tyr

530 535 540

Phe Cys Cys Cys Glu Gly Asn Met Cys Asn Glu Lys Phe Ser Tyr Phe

545 550 555 560

Pro Glu Met Glu Val Thr Gln Pro Thr Ser Gly Gly Gly Gly Ser Gly

565 570 575

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

580 585 590

Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp

595 600 605

Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys

610 615 620

Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys

625 630 635 640

Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val

645 650 655

Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr

660 665 670

Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp

675 680 685

Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu

690 695 700

Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile

705 710 715 720

Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

725 730

<210> 102

<211> 757

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A186

<400> 102

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ala Ile

450 455 460

Leu Gly Arg Ser Glu Thr Gln Glu Cys Leu Phe Tyr Asn Ala Asn Trp

465 470 475 480

Glu Leu Glu Arg Thr Asn Gln Thr Gly Val Glu Pro Cys Glu Gly Glu

485 490 495

Lys Asp Lys Arg Leu His Cys Tyr Ala Thr Trp Arg Asn Ile Ser Gly

500 505 510

Ser Ile Glu Ile Val Lys Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys

515 520 525

Tyr Asp Arg Thr Asp Cys Val Glu Thr Glu Glu Asn Pro Gln Val Tyr

530 535 540

Phe Cys Cys Cys Glu Gly Asn Met Cys Asn Glu Lys Phe Ser Tyr Phe

545 550 555 560

Pro Glu Met Glu Val Thr Gln Pro Thr Ser Gly Gly Gly Gly Ser Gly

565 570 575

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

580 585 590

Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met

595 600 605

Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala

610 615 620

His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn

625 630 635 640

Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe

645 650 655

Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr

660 665 670

Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys

675 680 685

Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu

690 695 700

Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile

705 710 715 720

Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys

725 730 735

Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn

740 745 750

Thr Ser Asn Pro Asp

755

<210> 103

<211> 734

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A187

<400> 103

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr

450 455 460

Gln Glu Cys Leu Phe Phe Asn Ala Asn Trp Glu Lys Asp Arg Thr Asn

465 470 475 480

Gln Thr Gly Val Glu Pro Cys Tyr Gly Asp Lys Asp Lys Arg Arg His

485 490 495

Cys Phe Ala Thr Trp Lys Asn Ile Ser Gly Ser Ile Glu Ile Val Lys

500 505 510

Gln Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Thr Asp Cys

515 520 525

Val Glu Lys Lys Asp Ser Pro Glu Val Tyr Phe Cys Cys Cys Glu Gly

530 535 540

Asn Met Cys Asn Glu Lys Phe Ser Tyr Phe Pro Glu Met Glu Val Thr

545 550 555 560

Gln Pro Thr Ser Asn Pro Val Thr Pro Lys Pro Pro Gly Gly Gly Gly

565 570 575

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

580 585 590

Gly Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn

595 600 605

Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln

610 615 620

Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys

625 630 635 640

Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln

645 650 655

Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu

660 665 670

Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu

675 680 685

Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro

690 695 700

Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp

705 710 715 720

Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

725 730

<210> 104

<211> 758

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A188

<400> 104

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Gln

450 455 460

Glu Cys Leu Phe Phe Asn Ala Asn Trp Glu Lys Asp Arg Thr Asn Gln

465 470 475 480

Thr Gly Val Glu Pro Cys Tyr Gly Asp Lys Asp Lys Arg Arg His Cys

485 490 495

Phe Ala Thr Trp Lys Asn Ile Ser Gly Ser Ile Glu Ile Val Lys Gln

500 505 510

Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Thr Asp Cys Val

515 520 525

Glu Lys Lys Asp Ser Pro Glu Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Met Cys Asn Glu Lys Phe Ser Tyr Phe Pro Glu Met Glu Val Thr Gln

545 550 555 560

Pro Thr Ser Asn Pro Val Thr Pro Lys Pro Pro Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

595 600 605

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

610 615 620

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

625 630 635 640

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

645 650 655

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

660 665 670

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

675 680 685

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

690 695 700

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

705 710 715 720

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

725 730 735

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

740 745 750

Asn Thr Ser Asn Pro Asp

755

<210> 105

<211> 733

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A189

<400> 105

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg Leu His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn

595 600 605

Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu

610 615 620

Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser

625 630 635 640

Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu

645 650 655

Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu

660 665 670

Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu

675 680 685

Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly

690 695 700

Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn

705 710 715 720

Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

725 730

<210> 106

<211> 758

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A190

<400> 106

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg Leu His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

595 600 605

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

610 615 620

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

625 630 635 640

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

645 650 655

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

660 665 670

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

675 680 685

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

690 695 700

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

705 710 715 720

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

725 730 735

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

740 745 750

Asn Thr Ser Asn Pro Asp

755

<210> 107

<211> 938

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A191

<400> 107

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Asn Cys

450 455 460

Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys Thr

465 470 475 480

Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr Ser

485 490 495

Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met Ile

500 505 510

Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys Glu

515 520 525

Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn Lys Lys Asn

530 535 540

Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp Lys

545 550 555 560

Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys Ala

565 570 575

Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln Tyr

580 585 590

Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly Ser

595 600 605

Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr Cys

610 615 620

Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys Gly

625 630 635 640

Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala Thr

645 650 655

Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys Ile

660 665 670

Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys Cys

675 680 685

Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp Glu

690 695 700

Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp Asn

705 710 715 720

Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser Ser

725 730 735

Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Ser Ile Ser

740 745 750

Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp Tyr Ser Phe

755 760 765

Pro Ile Ser Ser Ile Leu Glu Trp Gly Gly Gly Gly Ser Gly Gly Gly

770 775 780

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

785 790 795 800

Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp Met Ile

805 810 815

Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe

820 825 830

Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser

835 840 845

Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val

850 855 860

Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys

865 870 875 880

His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala

885 890 895

Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe

900 905 910

Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe

915 920 925

Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

930 935

<210> 108

<211> 963

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A192

<400> 108

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Asn Cys

450 455 460

Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys Thr

465 470 475 480

Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr Ser

485 490 495

Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met Ile

500 505 510

Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys Glu

515 520 525

Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn Lys Lys Asn

530 535 540

Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp Lys

545 550 555 560

Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys Ala

565 570 575

Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln Tyr

580 585 590

Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly Ser

595 600 605

Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr Cys

610 615 620

Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys Gly

625 630 635 640

Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala Thr

645 650 655

Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys Ile

660 665 670

Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys Cys

675 680 685

Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp Glu

690 695 700

Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp Asn

705 710 715 720

Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser Ser

725 730 735

Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Ser Ile Ser

740 745 750

Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp Tyr Ser Phe

755 760 765

Pro Ile Ser Ser Ile Leu Glu Trp Gly Gly Gly Gly Ser Gly Gly Gly

770 775 780

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

785 790 795 800

Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met Glu Ala

805 810 815

Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala His Pro

820 825 830

Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala

835 840 845

Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr

850 855 860

Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile

865 870 875 880

Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp

885 890 895

Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr

900 905 910

His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys

915 920 925

Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser

930 935 940

Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser

945 950 955 960

Asn Pro Asp

<210> 109

<211> 938

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A193

<400> 109

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Asn Cys

450 455 460

Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys Thr

465 470 475 480

Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr Ser

485 490 495

Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met Ile

500 505 510

Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys Glu

515 520 525

Asn Val Asp Cys Gly Pro Gly Gln Ser Cys Val Val Asp Gln Thr Gly

530 535 540

Ser Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp Lys

545 550 555 560

Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys Ala

565 570 575

Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln Tyr

580 585 590

Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly Ser

595 600 605

Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr Cys

610 615 620

Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys Gly

625 630 635 640

Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala Thr

645 650 655

Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys Ile

660 665 670

Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys Cys

675 680 685

Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp Glu

690 695 700

Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp Asn

705 710 715 720

Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser Ser

725 730 735

Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Ser Ile Ser

740 745 750

Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp Tyr Ser Phe

755 760 765

Pro Ile Ser Ser Ile Leu Glu Trp Gly Gly Gly Gly Ser Gly Gly Gly

770 775 780

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

785 790 795 800

Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp Met Ile

805 810 815

Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe

820 825 830

Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser

835 840 845

Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val

850 855 860

Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys

865 870 875 880

His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala

885 890 895

Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe

900 905 910

Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe

915 920 925

Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

930 935

<210> 110

<211> 963

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A194

<400> 110

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Asn Cys

450 455 460

Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys Thr

465 470 475 480

Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr Ser

485 490 495

Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met Ile

500 505 510

Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys Glu

515 520 525

Asn Val Asp Cys Gly Pro Gly Gln Ser Cys Val Val Asp Gln Thr Gly

530 535 540

Ser Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp Lys

545 550 555 560

Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys Ala

565 570 575

Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln Tyr

580 585 590

Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly Ser

595 600 605

Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr Cys

610 615 620

Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys Gly

625 630 635 640

Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala Thr

645 650 655

Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys Ile

660 665 670

Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys Cys

675 680 685

Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp Glu

690 695 700

Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp Asn

705 710 715 720

Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser Ser

725 730 735

Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Ser Ile Ser

740 745 750

Glu Asp Thr Glu Glu Glu Glu Glu Asp Glu Asp Gln Asp Tyr Ser Phe

755 760 765

Pro Ile Ser Ser Ile Leu Glu Trp Gly Gly Gly Gly Ser Gly Gly Gly

770 775 780

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

785 790 795 800

Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met Glu Ala

805 810 815

Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala His Pro

820 825 830

Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala

835 840 845

Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr

850 855 860

Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile

865 870 875 880

Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp

885 890 895

Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr

900 905 910

His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys

915 920 925

Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser

930 935 940

Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser

945 950 955 960

Asn Pro Asp

<210> 111

<211> 911

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A195

<400> 111

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Asn Cys

450 455 460

Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys Thr

465 470 475 480

Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr Ser

485 490 495

Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met Ile

500 505 510

Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys Glu

515 520 525

Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn Lys Lys Asn

530 535 540

Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp Lys

545 550 555 560

Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys Ala

565 570 575

Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln Tyr

580 585 590

Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly Ser

595 600 605

Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr Cys

610 615 620

Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys Gly

625 630 635 640

Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala Thr

645 650 655

Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys Ile

660 665 670

Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys Cys

675 680 685

Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp Glu

690 695 700

Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp Asn

705 710 715 720

Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser Ser

725 730 735

Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Gly Gly Gly

740 745 750

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

755 760 765

Ser Gly Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val

770 775 780

Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro

785 790 795 800

Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln

805 810 815

Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro

820 825 830

Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr

835 840 845

Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile

850 855 860

Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys

865 870 875 880

Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn

885 890 895

Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

900 905 910

<210> 112

<211> 936

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A196

<400> 112

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Asn Cys

450 455 460

Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys Thr

465 470 475 480

Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr Ser

485 490 495

Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met Ile

500 505 510

Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys Glu

515 520 525

Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn Lys Lys Asn

530 535 540

Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp Lys

545 550 555 560

Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys Ala

565 570 575

Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln Tyr

580 585 590

Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly Ser

595 600 605

Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr Cys

610 615 620

Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys Gly

625 630 635 640

Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala Thr

645 650 655

Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys Ile

660 665 670

Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys Cys

675 680 685

Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp Glu

690 695 700

Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp Asn

705 710 715 720

Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser Ser

725 730 735

Gly Val Leu Leu Glu Val Lys His Ser Gly Ser Cys Asn Gly Gly Gly

740 745 750

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

755 760 765

Ser Gly Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp

770 775 780

Val Glu Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn

785 790 795 800

Arg Thr Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr

805 810 815

Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp

820 825 830

Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys

835 840 845

Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val

850 855 860

Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp

865 870 875 880

Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro

885 890 895

Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met

900 905 910

Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu

915 920 925

Glu Tyr Asn Thr Ser Asn Pro Asp

930 935

<210> 113

<211> 733

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A197

<400> 113

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Glu Gly Asp Gln Asp Lys Arg Leu His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Lys Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn

595 600 605

Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu

610 615 620

Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser

625 630 635 640

Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu

645 650 655

Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu

660 665 670

Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu

675 680 685

Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly

690 695 700

Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn

705 710 715 720

Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

725 730

<210> 114

<211> 758

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A198

<400> 114

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Glu Gly Asp Gln Asp Lys Arg Leu His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Leu Asp Asp Ile Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Lys Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

595 600 605

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

610 615 620

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

625 630 635 640

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

645 650 655

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

660 665 670

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

675 680 685

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

690 695 700

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

705 710 715 720

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

725 730 735

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

740 745 750

Asn Thr Ser Asn Pro Asp

755

<210> 115

<211> 733

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A199

<400> 115

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Tyr Gly Asp Lys Asp Lys Arg Arg His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn

595 600 605

Asp Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu

610 615 620

Cys Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser

625 630 635 640

Cys Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu

645 650 655

Val Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu

660 665 670

Thr Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu

675 680 685

Asp Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly

690 695 700

Glu Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn

705 710 715 720

Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

725 730

<210> 116

<211> 758

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A200

<400> 116

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Tyr Gly Asp Lys Asp Lys Arg Arg His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Leu Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Ala Pro Thr Gly Gly Gly Gly Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

595 600 605

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

610 615 620

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

625 630 635 640

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

645 650 655

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

660 665 670

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

675 680 685

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

690 695 700

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

705 710 715 720

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

725 730 735

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

740 745 750

Asn Thr Ser Asn Pro Asp

755

<210> 117

<211> 730

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A201

<400> 117

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg Leu His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Asp Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Gly Gly Gly Gly Ser Gly Gly Gly

565 570 575

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

580 585 590

Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp Met Ile

595 600 605

Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe

610 615 620

Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser

625 630 635 640

Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val

645 650 655

Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys

660 665 670

His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala

675 680 685

Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe

690 695 700

Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe

705 710 715 720

Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

725 730

<210> 118

<211> 755

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A202

<400> 118

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Thr Arg

450 455 460

Glu Cys Ile Tyr Tyr Asn Ala Asn Trp Glu Leu Glu Arg Thr Asn Gln

465 470 475 480

Ser Gly Leu Glu Arg Cys Glu Gly Glu Gln Asp Lys Arg Leu His Cys

485 490 495

Tyr Ala Ser Trp Arg Asn Ser Ser Gly Thr Ile Glu Leu Val Lys Lys

500 505 510

Gly Cys Trp Asp Asp Asp Phe Asn Cys Tyr Asp Arg Gln Glu Cys Val

515 520 525

Ala Thr Glu Glu Asn Pro Gln Val Tyr Phe Cys Cys Cys Glu Gly Asn

530 535 540

Phe Cys Asn Glu Arg Phe Thr His Leu Pro Glu Ala Gly Gly Pro Glu

545 550 555 560

Val Thr Tyr Glu Pro Pro Pro Thr Gly Gly Gly Gly Ser Gly Gly Gly

565 570 575

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

580 585 590

Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met Glu Ala

595 600 605

Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala His Pro

610 615 620

Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn Gly Ala

625 630 635 640

Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe Ser Thr

645 650 655

Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr Ser Ile

660 665 670

Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys Asn Asp

675 680 685

Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu Pro Tyr

690 695 700

His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile Met Lys

705 710 715 720

Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys Ser Ser

725 730 735

Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn Thr Ser

740 745 750

Asn Pro Asp

755

<210> 119

<211> 732

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A203

<400> 119

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ala Ile

450 455 460

Leu Gly Arg Ser Glu Thr Gln Glu Cys Leu Phe Tyr Asn Ala Asn Trp

465 470 475 480

Glu Leu Glu Arg Thr Asn Gln Thr Gly Val Glu Pro Cys Glu Gly Glu

485 490 495

Lys Asp Lys Arg Leu His Cys Tyr Ala Thr Trp Arg Asn Ile Ser Gly

500 505 510

Ser Ile Glu Ile Val Lys Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys

515 520 525

Tyr Asp Arg Thr Asp Cys Val Glu Thr Glu Glu Asn Pro Gln Val Tyr

530 535 540

Phe Cys Cys Cys Glu Gly Asn Met Cys Asn Glu Lys Phe Ser Tyr Phe

545 550 555 560

Pro Glu Met Glu Val Thr Gln Pro Thr Ser Gly Gly Gly Gly Ser Gly

565 570 575

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

580 585 590

Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Val Asn Asn Asp

595 600 605

Met Ile Val Thr Asp Asn Asn Gly Ala Val Lys Phe Pro Gln Leu Cys

610 615 620

Lys Phe Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys

625 630 635 640

Met Ser Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val

645 650 655

Cys Val Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr

660 665 670

Val Cys His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp

675 680 685

Ala Ala Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu

690 695 700

Thr Phe Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile

705 710 715 720

Ile Phe Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp

725 730

<210> 120

<211> 757

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A204

<400> 120

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ala Ile

450 455 460

Leu Gly Arg Ser Glu Thr Gln Glu Cys Leu Phe Tyr Asn Ala Asn Trp

465 470 475 480

Glu Leu Glu Arg Thr Asn Gln Thr Gly Val Glu Pro Cys Glu Gly Glu

485 490 495

Lys Asp Lys Arg Leu His Cys Tyr Ala Thr Trp Arg Asn Ile Ser Gly

500 505 510

Ser Ile Glu Ile Val Lys Lys Gly Cys Trp Leu Asp Asp Phe Asn Cys

515 520 525

Tyr Asp Arg Thr Asp Cys Val Glu Thr Glu Glu Asn Pro Gln Val Tyr

530 535 540

Phe Cys Cys Cys Glu Gly Asn Met Cys Asn Glu Lys Phe Ser Tyr Phe

545 550 555 560

Pro Glu Met Glu Val Thr Gln Pro Thr Ser Gly Gly Gly Gly Ser Gly

565 570 575

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

580 585 590

Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu Met

595 600 605

Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr Ala

610 615 620

His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn Asn

625 630 635 640

Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg Phe

645 650 655

Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile Thr

660 665 670

Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg Lys

675 680 685

Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys Leu

690 695 700

Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys Ile

705 710 715 720

Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser Cys

725 730 735

Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr Asn

740 745 750

Thr Ser Asn Pro Asp

755

<210> 121

<211> 534

<212> PRT

<213> Artificial

<220>

<223> Amino acid sequence A216

<400> 121

Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala

1 5 10 15

Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

20 25 30

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

35 40 45

Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val

50 55 60

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

65 70 75 80

Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

85 90 95

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala

100 105 110

Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

115 120 125

Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr

130 135 140

Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser

145 150 155 160

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

165 170 175

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

180 185 190

Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe

195 200 205

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

210 215 220

Ser Leu Ser Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly

225 230 235 240

Gly Ser Gly Gly Gly Gly Ser Glu Thr Gln Glu Cys Leu Phe Phe Asn

245 250 255

Ala Asn Trp Glu Lys Asp Arg Thr Asn Gln Thr Gly Val Glu Pro Cys

260 265 270

Tyr Gly Asp Lys Asp Lys Arg Arg His Cys Phe Ala Thr Trp Lys Asn

275 280 285

Ile Ser Gly Ser Ile Glu Ile Val Lys Gln Gly Cys Trp Leu Asp Asp

290 295 300

Ile Asn Cys Tyr Asp Arg Thr Asp Cys Val Glu Lys Lys Asp Ser Pro

305 310 315 320

Glu Val Tyr Phe Cys Cys Cys Glu Gly Asn Met Cys Asn Glu Lys Phe

325 330 335

Ser Tyr Phe Pro Glu Met Glu Val Thr Gln Pro Thr Ser Asn Pro Val

340 345 350

Thr Pro Lys Pro Pro Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

355 360 365

Gly Gly Gly Ser Thr Ile Pro Pro His Val Gln Lys Ser Asp Val Glu

370 375 380

Met Glu Ala Gln Lys Asp Glu Ile Ile Cys Pro Ser Cys Asn Arg Thr

385 390 395 400

Ala His Pro Leu Arg His Ile Asn Asn Asp Met Ile Val Thr Asp Asn

405 410 415

Asn Gly Ala Val Lys Phe Pro Gln Leu Cys Lys Phe Cys Asp Val Arg

420 425 430

Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser Asn Cys Ser Ile

435 440 445

Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val Ala Val Trp Arg

450 455 460

Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys His Asp Pro Lys

465 470 475 480

Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala Ser Pro Lys Cys

485 490 495

Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe Phe Met Cys Ser

500 505 510

Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe Ser Glu Glu Tyr

515 520 525

Asn Thr Ser Asn Pro Asp

530

<210> 122

<211> 584

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the heavy chain of A167

<400> 122

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Gly Leu Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ile Pro Tyr Asn Gly Asn Thr Asn Ser Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Arg Asp Arg Asp Tyr Gly Val Asn Tyr Asp Ala Phe Asp Ile Trp

100 105 110

Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Val Gln Ser

130 135 140

Gly Ala Glu Val Lys Lys Pro Gly Ser Ser Val Lys Val Ser Cys Lys

145 150 155 160

Ala Ser Gly Tyr Thr Phe Ser Ser Asn Val Ile Ser Trp Val Arg Gln

165 170 175

Ala Pro Gly Gln Gly Leu Glu Trp Met Gly Gly Val Ile Pro Ile Val

180 185 190

Asp Ile Ala Asn Tyr Ala Gln Arg Phe Lys Gly Arg Val Thr Ile Thr

195 200 205

Ala Asp Glu Ser Thr Ser Thr Thr Tyr Met Glu Leu Ser Ser Leu Arg

210 215 220

Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Ser Thr Leu Gly Leu Val

225 230 235 240

Leu Asp Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

245 250 255

Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser

260 265 270

Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp

275 280 285

Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr

290 295 300

Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr

305 310 315 320

Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys

325 330 335

Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp

340 345 350

Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala

355 360 365

Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

370 375 380

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

385 390 395 400

Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val

405 410 415

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

420 425 430

Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

435 440 445

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly

450 455 460

Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

465 470 475 480

Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr

485 490 495

Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser

500 505 510

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

515 520 525

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

530 535 540

Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe

545 550 555 560

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

565 570 575

Ser Leu Ser Leu Ser Leu Gly Lys

580

<210> 123

<211> 337

<212> PRT

<213> Artificial

<220>

<223>Amino acid sequence of the light chain of A167

<400> 123

Ser Tyr Glu Val Thr Gln Ala Pro Ser Val Ser Val Ser Pro Gly Gln

1 5 10 15

Thr Ala Ser Ile Thr Cys Ser Gly Asp Lys Leu Gly Asp Lys Tyr Ala

20 25 30

Cys Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Val Leu Val Ile Tyr

35 40 45

Gln Asp Ser Lys Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser

50 55 60

Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Met

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Trp Asp Ser Ser Thr Ala Val

85 90 95

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly Ser Gly

100 105 110

Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Thr Val Leu Thr Gln Ser

115 120 125

Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys

130 135 140

Arg Ala Ser Gln Ser Leu Gly Ser Ser Tyr Leu Ala Trp Tyr Gln Gln

145 150 155 160

Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg

165 170 175

Ala Pro Gly Ile Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp

180 185 190

Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr

195 200 205

Tyr Cys Gln Gln Tyr Ala Asp Ser Pro Ile Thr Phe Gly Gln Gly Thr

210 215 220

Arg Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe

225 230 235 240

Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys

245 250 255

Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val

260 265 270

Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln

275 280 285

Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser

290 295 300

Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His

305 310 315 320

Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

325 330 335

Ala

<210> 124

<211> 12

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 124

Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser

1 5 10

<210> 125

<211> 4

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 125

Gly Gly Gly Ser

1

<210> 126

<211> 13

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 126

Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly

1 5 10

<210> 127

<211> 5

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 127

Gly Ser Ser Gly Thr

1 5

<210> 128

<211> 14

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 128

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Ser

1 5 10

<210> 129

<211> 17

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 129

Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys

1 5 10 15

Ala

<210> 130

<211> 20

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 130

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

1 5 10 15

Gly Gly Gly Ser

20

<210> 131

<211> 8

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 131

Gly Gly Gly Ser Gly Gly Gly Ser

1 5

<210> 132

<211> 5

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 132

Gly Ser Gly Ser Thr

1 5

<210> 133

<211> 4

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 133

Gly Gly Ser Ser

1

<210> 134

<211> 5

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 134

Gly Gly Gly Gly Ser

1 5

<210> 135

<211> 4

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 135

Gly Gly Ser Gly

1

<210> 136

<211> 4

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 136

Ser Gly Gly Gly

1

<210> 137

<211> 4

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 137

Gly Ser Gly Ser

1

<210> 138

<211> 6

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 138

Gly Ser Gly Ser Gly Ser

1 5

<210> 139

<211> 8

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 139

Gly Ser Gly Ser Gly Ser Gly Ser

1 5

<210> 140

<211> 10

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 140

Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser

1 5 10

<210> 141

<211> 12

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 141

Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser

1 5 10

<210> 142

<211> 10

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 142

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

1 5 10

<210> 143

<211> 15

<212> PRT

<213> Artificial

<220>

<223> Peptide linker sequence

<400> 143

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

1 5 10 15

<---

Claims (6)

1. An isolated multispecific antagonist molecule comprising a first antigen-binding molecule that specifically binds to activin or an activin receptor and a second antigen-binding molecule that specifically binds to a TGF-β receptor, wherein said multispecific antagonist molecule simultaneously neutralizes the activin signaling pathway and the TGF-β signaling pathway, wherein said multispecific antagonist molecule comprises an amino acid sequence selected from the group consisting of the amino acid sequences presented in SEQ ID NO: 54, 55, 60 and 61. 2. An isolated multispecific antagonist molecule according to claim 1, wherein the molecule of said multispecific antagonist comprises the amino acid sequence presented in SEQ ID NO: 55. 3. An isolated multispecific antagonist molecule comprising a first antigen-binding molecule that specifically binds to activin or an activin receptor and a second antigen-binding molecule that specifically binds to a TGF-β receptor, wherein said multispecific antagonist molecule simultaneously neutralizes the activin signaling pathway and the TGF-β signaling pathway, wherein the activin-binding polypeptide is an antibody comprising the heavy chain variable region sequence presented in SEQ ID NO: 12 and the light chain variable region sequence presented in SEQ ID NO: 13; and wherein the TGF-β-binding polypeptide comprises SEQ ID NO: 21. 4. An isolated multispecific antagonist molecule comprising a first antigen-binding molecule that specifically binds to activin or an activin receptor and a second antigen-binding molecule that specifically binds to a TGF-β receptor, wherein said multispecific antagonist molecule simultaneously neutralizes the activin signaling pathway and the TGF-β signaling pathway, wherein the activin-binding polypeptide is an antibody comprising the heavy chain variable region sequence presented in SEQ ID NO: 12 and the light chain variable region sequence presented in SEQ ID NO: 13; and wherein the TGF-β-binding polypeptide is an antibody comprising the heavy chain variable region sequence presented in SEQ ID NO: 24 and the light chain variable region sequence presented in SEQ ID NO: 25. 5. A pharmaceutical composition that simultaneously neutralizes the activin signaling pathway and the TGF-β signaling pathway, wherein said composition contains a therapeutically effective amount of a multispecific antagonist molecule according to any one of claims 1-4 in a mixture with a pharmaceutically acceptable carrier. 6. The pharmaceutical composition of claim 5, for use in the treatment of cancer, wherein the cancer is selected from the group consisting of melanoma, multiple myeloma, lung cancer, pancreatic cancer, colorectal cancer, liver cancer, stomach cancer, kidney cancer, bladder cancer, head and neck cancer, thyroid cancer, breast cancer, ovarian cancer, endometrial cancer, testicular cancer, prostate cancer, and brain cancer.