CN115843256A - anti-ERBB 3 antibody or antigen binding fragment thereof and medical application thereof - Google Patents

Abstract

Relates to an anti-ERBB 3 antibody or an antigen binding fragment thereof and medical application thereof, in particular to a fully humanized antibody or an antigen binding fragment thereof for resisting an ERBB3 receptor and medical application thereof. Also comprises anti-ERBB 3 fully human antibody or antigen binding fragment thereof, and application thereof as an anti-cancer drug.

Description

anti-ERBB 3 antibody or antigen binding fragment thereof and medical application thereof Technical Field

The present invention relates to an ERBB3 antibody or an antigen-binding fragment thereof, and further, the present invention relates to an ERBB3 fully human antibody or an antigen-binding fragment thereof comprising a CDR region; the invention also relates to a pharmaceutical composition containing the ERBB3 fully human antibody or the antigen binding fragment thereof, and application of the pharmaceutical composition as a diagnostic agent and a therapeutic drug for the ERBB3 related diseases.

Background

ERBB receptors are widely expressed in neuronal, epithelial, mesenchymal cells, involved in the development of cardiovascular, neural, musculoskeletal and other organs, and involved in cancer pathogenesis. There are 4 family proteins for ERBB receiver: EGFR, ERBB2, ERBB3, and ERBB4, which are membrane proteins, have similar molecular structures, and comprise an extracellular region (ECD), a transmembrane region, an intracellular region having tyrosine kinase activity, and a C-terminal. ECD can be subdivided into 4 subdomains: subdomain I, II, III, IV (BMC biolnformatics 2001, 2; mol Cell 2003, 11; II and IV are cysteine-rich and are functional domains that form dimers. In addition to ERBB2, three ERBB receptors bind II and IV in an inactive state and only open when bound and activated by a ligand, leaving II exposed to form a dimer. At present, more EGFR and ERBB2 are researched, and the corresponding targeted drugs are more intensively developed. In recent years, the interest of ERBB3 is increasing, and ERBB3 bypass plays a key role in EGFR and ERBB2 related drug resistance, and ERBB3 has high expression in tumors such as breast cancer, lung cancer, prostate cancer, colorectal cancer, ovarian cancer, gastric cancer, bladder cancer, melanoma, and the like, so ERBB3 is another potential target for tumor therapy. ERBB3 has 2 ligands NRG1 and NRG2, the ligands can activate the kinase activity of ERBB3, and the activated ERBB3 can form heterodimer with EGFR or ERBB2 and phosphorylate the latter, so that signals are transmitted to a downstream pathway to promote the growth and proliferation of tumor cells. In addition, ERBB3 may also signal by "cross talk" with other RTKs in cancer, such as the formation of complexes with IGF1R, FGFR and HGFR (c-Met) (BioDrugs 2017,31, 63-73), which shows that ERBB3 plays an important role in tumor growth.

Currently, therapeutic strategies against ERBB3 targets are based on several mechanisms: 1) Locking ERBB3 in an inactive state (e.g., CDX-3379); 2) Ligand NRG (e.g. RB 200) to capture ERBB 3; 3) Block binding of ERBB3 to a ligand (e.g., U3-1287); 4) Promoting the endocytosis of ERBB3 (antibody drug conjugate); 5) Blocking dimerization of ERBB3 with other EGFR family members; 6) Recruiting immune cells to kill cancer cells expressing endogenous ERBB3. . For these mechanisms, targeted therapies for ERBB3 may be: monoclonal antibodies, bispecific antibodies, vaccines against ERBB3, ligand trap, RNA inhibitors that lock onto ERBB3, small molecules that inhibit ERBB3 kinase activity, and the like. Among them, the developing heat of the antibody targeting ERBB3 is high.

In clinical studies, an ERBB3 antibody from murine origin significantly inhibited the growth of patient cytoma (data from Celldex are cited). The ERBB3 antibody which is currently researched is a humanized antibody obtained by performing humanized transformation on a murine antibody, and the humanized antibody has higher immunogenicity in immunization than a fully humanized antibody without a murine antibody component, and is an adverse factor in human application; some of the ERBB3 antibody drugs under investigation are fully human antibodies obtained by phage display technology, which can solve the problem of immunogenicity.

The phage display technology (phage display technology) expresses a foreign protein or polypeptide by fusion with a phage coat protein, thereby expressing the foreign protein on the surface of a phage. The phage antibody library is an antibody library established by combining a phage display technology, a PCR amplification technology and a protein expression technology by using a comprehensive technical means.

The phage library generally comprises a synthetic library, an immune library and a natural library, the natural library is used more frequently, the natural library is a fully human antibody library constructed by immune cells of human peripheral blood, and the greatest advantage is that the fully human antibody with high diversity generated by a human body can be obtained without additional in vivo immunity. In addition, the phage antibody library has the following advantages: (1) the unification of genotype and phenotype is realized. In addition, the experimental method is simple and rapid, the traditional antibody production method by the hybridoma technology needs to take several months, and the antibody library technology only needs to be short for several weeks. (2) The expressed antibody is completely humanized antibody, has small molecular weight, is mainly expressed in the form of active fragment Fab and scFV, and has obvious advantage in the aspect of tissue penetration compared with the intact antibody. (3) The screening capacity is large, the hybridoma technology is screened in thousands of clones, and the antibody library technology can select millions or even hundreds of millions of molecules. The antibodies screened were more diverse. (4) The application is wide, a prokaryotic expression system is adopted, and the advantages are more obvious when the method is used for large-scale production (Curr Opin Biotechnology.2002 Dec;13 (6): 598-602.

At present, patents such as WO2007077028 report antibodies to ERBB3, most of which are murine antibodies or humanized antibodies, and some of which are fully humanized antibodies obtained from phage libraries, most of which are in the early clinical stage and discovery stage at home and abroad, no antibody medicine targeting ERBB3 is available on the market, so that further development of ERBB3 fully humanized antibodies with higher activity, high affinity and high stability is necessary for therapeutic research and application of related diseases.

Disclosure of Invention

The invention provides an anti-ERBB 3 antibody or an antigen-binding fragment thereof, and particularly relates to a fully human anti-ERBB 3 antibody or an antigen-binding fragment thereof, wherein the anti-ERBB 3 antibody or the antigen-binding fragment thereof comprises: antibody heavy chain variable region and light chain variable region; wherein the variable region of the antibody heavy chain comprises at least 1 HCDR selected from the group consisting of SEQ ID NOs: 1,2, 3 or 4; the variable region of the antibody light chain comprises at least 1 LCDR selected from the group consisting of SEQ ID NOs: 5, 6 or 7.

The present invention also relates to a preferred embodiment, an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, wherein:

the heavy chain variable region of the antibody comprises HCDR1 shown in SEQ ID NO.1, HCDR2 shown in SEQ ID NO.2 and HCDR3 shown in SEQ ID NO. 4; or the like, or, alternatively,

the heavy chain variable region of the antibody comprises HCDR1 shown in SEQ ID NO.1, HCDR2 shown in SEQ ID NO. 3 and HCDR3 shown in SEQ ID NO. 4.

The present invention also relates to a preferred embodiment, an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, wherein said antibody light chain variable region comprises:

LCDR1 shown in SEQ ID NO. 5, LCDR2 shown in SEQ ID NO. 6, and LCDR3 shown in SEQ ID NO. 7.

The present invention also relates to a preferred embodiment, an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, which comprises a heavy chain variable region and a light chain variable region, wherein:

wherein the variable region of the antibody heavy chain comprises HCDR1 shown in SEQ ID NO.1, HCDR2 shown in SEQ ID NO.2 and HCDR3 shown in SEQ ID NO. 4; the variable region of the antibody light chain comprises LCDR1 shown in SEQ ID NO. 5, LCDR2 shown in SEQ ID NO. 6 and LCDR3 shown in SEQ ID NO. 7.

The present invention also relates to a preferred embodiment, an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, which comprises a heavy chain variable region and a light chain variable region, wherein:

the variable region of the antibody heavy chain comprises: HCDR1 shown in SEQ ID NO.1, HCDR2 shown in SEQ ID NO. 3, and HCDR3 shown in SEQ ID NO. 4; the antibody light chain variable region comprises: LCDR1 shown in SEQ ID NO. 5, LCDR2 shown in SEQ ID NO. 6, and LCDR3 shown in SEQ ID NO. 7.

The present invention also relates to a preferred embodiment, an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, wherein the heavy chain variable region of said antibody is a heavy chain variable region selected from the group consisting of those represented by the following sequences: SEQ ID NO 8 or SEQ ID NO 9.

The present invention also relates to a preferred embodiment, an anti-ERBB 3 antibody or an antigen-binding fragment thereof as described above, wherein the variable region of the light chain of the antibody is SEQ ID NO. 10.

The present invention also relates to a preferred embodiment, an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, wherein said antibody comprises: the heavy chain variable region shown by SEQ ID NO. 8 and the light chain variable region shown by SEQ ID NO. 10; or the like, or, alternatively,

the anti-ERBB 3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region shown in SEQ ID NO. 9 and a light chain variable region shown in SEQ ID NO. 10.

In a preferred embodiment of the invention, the anti-ERBB 3 antibody or antigen-binding fragment thereof further comprises a heavy chain constant region comprising a heavy chain derived from human IgG1, igG2, igG3, or IgG4, or a mutant thereof;

in a further preferred embodiment of the invention, the anti-ERBB 3 antibody or antigen-binding fragment thereof further comprises a heavy chain constant region of human IgG1 or a variant thereof;

in a further preferred embodiment of the invention, the anti-ERBB 3 antibody or antigen-binding fragment thereof further comprises a human IgG1 heavy chain constant region;

in a further preferred embodiment of the invention, the anti-ERBB 3 antibody or antigen-binding fragment thereof further comprises a heavy chain constant region as shown in SEQ ID NO. 17;

in a further preferred embodiment of the invention, the anti-ERBB 3 antibody or antigen-binding fragment thereof further comprises a light chain constant region derived from a human kappa chain, lambda chain, or mutant thereof;

in a further preferred embodiment of the invention, the anti-ERBB 3 antibody or antigen-binding fragment thereof further comprises a light chain constant region derived from a human λ chain.

In a further preferred embodiment of the invention, the anti-ERBB 3 antibody or antigen-binding fragment thereof further comprises a light chain constant region as shown in SEQ ID NO. 18.

In one preferred embodiment of the invention, the anti-ERBB 3 antibody or antigen-binding fragment thereof, wherein said anti-ERBB 3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region selected from the group consisting of: 8, 9, or a heavy chain variable region having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% homology thereto.

In a preferred embodiment of the invention, the anti-ERBB 3 antibody or antigen-binding fragment thereof, wherein the anti-ERBB 3 antibody or antigen-binding fragment thereof comprises the light chain variable region SEQ ID NO:10, or a light chain variable region having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% homology thereto.

The present invention also relates to a preferred embodiment, an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, wherein the heavy chain of said antibody is selected from the group consisting of the heavy chains having the following sequences: SEQ ID NO 11 or SEQ ID NO 12.

The present invention also relates to a preferred embodiment, an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, wherein the light chain of the antibody is SEQ ID NO. 13.

The invention also relates to a preferred embodiment, an anti-ERBB 3 antibody or an antigen-binding fragment thereof as described above, wherein,

the anti-ERBB 3 antibody comprises a heavy chain shown as SEQ ID NO.11 and a light chain shown as SEQ ID NO. 13; or the like, or, alternatively,

the anti-ERBB 3 antibody comprises a heavy chain shown as SEQ ID NO. 12 and a light chain shown as SEQ ID NO. 13.

The present invention also relates to a preferred embodiment, an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, wherein said anti-ERBB 3 antibody or antigen-binding fragment thereof comprises a heavy chain selected from the group consisting of: SEQ ID NO: 11. 12, or a full-length heavy chain having at least 80%, 85%, 90%, 95%, or 99% homology thereto.

The present invention also relates to a preferred embodiment, an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, wherein said anti-ERBB 3 antibody or antigen-binding fragment thereof comprises a light chain of SEQ ID NO:13, or a full length light chain having at least 80%, 85%, 90%, 95%, or 99% homology thereto.

The invention further provides a polynucleotide encoding the anti-ERBB 3 antibody or antigen-binding fragment thereof described above.

The present invention further provides an expression vector comprising the polynucleotide described above.

The present invention further provides a host cell into which the above-described expression vector is introduced or contained.

In a preferred embodiment of the present invention, the host cell is a bacterium, preferably E.coli.

In a preferred embodiment of the present invention, the host cell is a yeast, preferably pichia pastoris.

In a preferred embodiment of the invention, the above-mentioned host cell is a mammalian cell, preferably a CHO cell or a HEK293 cell.

The present invention further provides a method of producing an anti-ERBB 3 antibody or antigen-binding fragment thereof, comprising the steps of:

a) Culturing the above host cell;

b) Isolating fully human antibodies from the culture; and

c) Purifying the antibody.

The invention further provides a pharmaceutical composition comprising an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, and a pharmaceutically acceptable excipient, diluent, or carrier.

The invention further provides a detection or diagnostic reagent comprising an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, and an excipient, diluent, or carrier useful for detection or diagnosis.

The invention further provides the use of an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, or a composition as described above, in the manufacture of a medicament for the treatment or prevention of an ERBB3 mediated disease or disorder.

The invention further provides the use of an anti-ERBB 3 antibody or antigen-binding fragment thereof as described above, or a detection or diagnostic reagent as described above, in the preparation of a reagent for detecting, diagnosing, prognosing an ERBB3 mediated disease or disorder.

In a preferred embodiment of the present invention,

the disease or disorder described above is cancer;

preferably a cancer in which ERBB3 is expressed or overexpressed;

more preferably breast cancer, ovarian cancer, prostate cancer, endometrial cancer, thyroid cancer, renal cancer, lung cancer, gastric cancer, colon cancer, bladder cancer, cervical cancer, gallbladder cancer, pancreatic cancer, testicular cancer, soft tissue sarcoma, head and neck cancer, glioma or melanoma.

In a preferred embodiment of the invention, the above-described anti-ERBB 3 antibody or antigen-binding fragment thereof, or the above-described composition, is used for the detection, diagnosis, prognosis of ERBB 3-mediated diseases; the disease is selected from: breast cancer, ovarian cancer, prostate cancer, endometrial cancer, thyroid cancer, renal cancer, lung cancer, gastric cancer, colon cancer, bladder cancer, cervical cancer, gallbladder cancer, pancreatic cancer, testicular cancer, soft tissue sarcoma, head and neck cancer, glioma, or melanoma.

The present invention further provides a method of treating or preventing an ERBB3 mediated disease, comprising the steps of:

providing to the subject a therapeutically effective amount or a prophylactically effective amount of the anti-ERBB 3 antibody or antigen-binding fragment thereof described above; or providing a therapeutically effective amount or a prophylactically effective amount of the pharmaceutical composition described above to a subject; wherein the ERBB3 mediated disease is selected from: breast cancer, ovarian cancer, prostate cancer, endometrial cancer, thyroid cancer, renal cancer, lung cancer, gastric cancer, colon cancer, bladder cancer, cervical cancer, gallbladder cancer, pancreatic cancer, testicular cancer, soft tissue sarcoma, head and neck cancer, glioma, or melanoma.

Detailed Description

Detailed Description

1. Term(s) for

In order that the invention may be more readily understood, certain technical and scientific terms are specifically defined below. Unless clearly defined otherwise herein, all other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

The three letter codes and the one letter codes for amino acids used in the present invention are as described in j.biol.chem,243, p3558 (1968).

The term "antibody" as used herein refers to an immunoglobulin, which is a tetrapeptide chain structure formed by two identical heavy chains and two identical light chains linked by interchain disulfide bonds. The constant regions of immunoglobulin heavy chains differ in their amino acid composition and arrangement, and thus, their antigenicity. Accordingly, immunoglobulins can be classified into five classes, otherwise known as the immunoglobulin isotypes, namely, igM, igD, igG, igA, and IgE, with their corresponding heavy chains being the μ chain, the δ chain, the γ chain, the α chain, and the ε chain, respectively. The same class of igs can be divided into different subclasses according to differences in amino acid composition of the hinge region and the number and position of disulfide bonds in the heavy chain, and for example, iggs can be classified into IgG1, igG2, igG3 and IgG4. Light chains are classified as either kappa or lambda chains by differences in the constant regions. The second of the five classes of Ig may have either a kappa chain or a lambda chain.

The antibody light chain variable region of the invention may further comprise a light chain constant region comprising a human or murine kappa, lambda chain or variant thereof.

The antibody heavy chain variable region of the invention may further comprise a heavy chain constant region comprising human or murine IgG1, igG2, igG3, igG4 or variants thereof.

The sequences of the antibody heavy and light chains, near the N-terminus, are widely varied by about 110 amino acids, being variable regions (V-regions); the remaining amino acid sequence near the C-terminus is relatively stable and is a constant region (C-region). The variable regions include 3 hypervariable regions (HVRs) and 4 Framework Regions (FRs) which are relatively sequence-conserved. The 3 hypervariable regions determine the specificity of the antibody, and are also known as Complementarity Determining Regions (CDRs). Each of the light chain variable region (VL) and the heavy chain variable region (VH) is composed of 3 CDR regions and 4 FR regions, and the sequence from the amino terminus to the carboxyl terminus is: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The 3 CDR regions of the light chain refer to LCDR1, LCDR2, and LCDR3; the 3 CDR regions of the heavy chain refer to HCDR1, HCDR2 and HCDR3. The CDR amino acid residues in the VL and VH regions of the antibodies or antigen-binding fragments of the invention conform in number and position to the rules of the known Kabat or Chothia or ABM definitions (http:// bio in. Org. Uk/abs /).

The term "antigen presenting cell" or "APC" is a cell that displays foreign antigens complexed with MHC on its surface. T cells recognize this complex using the T Cell Receptor (TCR). Examples of APCs include, but are not limited to, dendritic Cells (DCs), peripheral Blood Mononuclear Cells (PBMCs), monocytes, B lymphoblastoid cells, and monocyte-derived dendritic cells.

The term "antigen presentation" refers to the process by which APCs capture antigens and enable them to be recognized by T cells, for example as a component of an MHC-I/MHC-II conjugate.

The term "recombinant human antibody" includes human antibodies made, expressed, created or isolated by recombinant methods, involving techniques and methods well known in the art, such as:

1. antibodies isolated from transgenic, transchromosomal animals (e.g., mice) of human immunoglobulin genes or hybridomas prepared therefrom;

2. antibodies isolated from host cells transformed to express the antibodies, such as transfectomas;

3. antibodies isolated from a library of recombinant combinatorial human antibodies; and

4. antibodies produced, expressed, created or isolated by methods such as splicing of human immunoglobulin gene sequences to other DNA sequences.

Such recombinant human antibodies comprise variable and constant regions that utilize specific human germline immunoglobulin sequences encoded by germline genes, but also include subsequent rearrangements and mutations such as occur during antibody maturation.

The term "human antibody" includes antibodies having variable and constant regions of human germline immunoglobulin sequences. The human antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, the term "human antibody" does not include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences (i.e., "humanized antibodies").

The term "antigen-binding fragment" refers to antigen-binding fragments and antibody analogs of antibodies, which typically include at least a portion of the antigen-binding or variable region (e.g., one or more CDRs) of a parent antibody. Antibody fragments retain at least some of the binding specificity of the parent antibody. Typically, an antibody fragment retains at least 10% of parent binding activity when expressed as activity on a molar basis. Preferably, the antibody fragment retains at least 20%, 50%, 70%, 80%, 90%, 95%, or 100% or more of the binding affinity of the parent antibody to the target. Examples of antigen-binding fragments include, but are not limited to: fab, fab ', F (ab') 2, fv fragments, linear antibodies, single chain antibodies, nanobodies, domain antibodies and multispecific antibodies. Engineered antibody variants are reviewed in Holliger and Hudson,2005, nat. Biotechnol.23: 1126-1136.

The term "Fab fragment" consists of the CH1 and variable regions of one light and one heavy chain. The heavy chain of a Fab molecule cannot form a disulfide bond with another heavy chain molecule.

The term "Fc" region contains two heavy chain fragments comprising the CH1 and CH2 domains of an antibody. The two heavy chain fragments are held together by two or more disulfide bonds and by the hydrophobic interaction of the CH3 domains.

The term "Fab ' fragment" contains a portion of one light chain and one heavy chain comprising the VH domain and the CH1 domain and the region between the CH1 and CH2 domains, whereby an interchain disulfide bond can be formed between the two heavy chains of the two Fab ' fragments to form the F (ab ') 2 molecule.

The term "F (ab') 2 fragment" contains two light chains and two heavy chains comprising part of the constant region between the CH1 and CH2 domains, whereby an interchain disulfide bond is formed between the two heavy chains. Thus, a F (ab ') 2 fragment consists of two Fab' fragments held together by a disulfide bond between the two heavy chains.

The term "Fv region" comprises variable regions from both the heavy and light chains, but lacks a constant region.

The term "multispecific antibody" is used in its broadest sense to encompass antibodies having polyepitopic specificity. These multispecific antibodies include, but are not limited to: an antibody comprising a heavy chain variable region VH and a light chain variable region VL, wherein the VH-VL unit has polyepitopic specificity; an antibody having two or more VL and VH regions, each VH-VL unit binding to a different target or a different epitope of the same target; an antibody having two or more single variable regions, each single variable region binding to a different target or a different epitope of the same target; full length antibodies, antibody fragments, diabodies (diabodies), bispecific diabodies and triabodies (triabodies), antibody fragments that have been covalently or non-covalently linked together, and the like.

The term "single-chain antibody" is a single-chain recombinant protein formed by connecting a heavy chain variable region VH and a light chain variable region VL of an antibody via a linker peptide, and is the smallest antibody fragment having a complete antigen-binding site.

The term "domain antibody fragment" is an immunologically functional immunoglobulin fragment that contains only heavy chain variable regions or light chain variable regions. In certain instances, two or more VH regions are covalently linked to a peptide linker to form a bivalent domain antibody fragment. The two VH regions of the bivalent domain antibody fragment may target the same or different antigens.

The term "binds to ERBB 3" as used herein refers to a molecule that interacts with human ERBB3.

The term "antigen binding site" refers to a three-dimensional spatial site recognized by an antibody or antigen binding fragment of the invention.

The term "epitope" refers to a site on an antigen to which an immunoglobulin or antibody specifically binds. Epitopes can be formed from contiguous amino acids, or non-contiguous amino acids juxtaposed by tertiary folding of the protein. Epitopes formed by adjacent amino acids are typically retained after exposure to denaturing solvents, whereas epitopes formed by tertiary folding are typically lost after denaturing solvent treatment. Epitopes typically comprise at least 3-15 amino acids in a unique spatial conformation. Methods of determining what epitope is bound by a given antibody are well known in the art and include immunoblot and immunoprecipitation detection assays, and the like. Methods of determining the spatial conformation of an epitope include techniques in the art and techniques described herein, such as X-ray crystallography and two-dimensional nuclear magnetic resonance, among others.

The terms "specific binding", "selective binding" and "selective binding" refer to the binding of an antibody to an epitope on a predetermined antigen. Typically, antibodies are less than about 10 angstroms when measured in an instrument by Surface Plasmon Resonance (SPR) techniques using human ERBB3 as the analyte and antibodies as the ligand ^-7 M or even smaller equilibrium dissociation constant (K) _D ) Binds to a predetermined antigen with an affinity for the predetermined antigenIt binds with at least twice the affinity to a non-specific antigen other than the predetermined antigen or closely related antigens (e.g., BSA, etc.). The term "antibody recognizing an antigen" is used interchangeably herein with the term "specifically binding antibody".

The term "cross-reactive" refers to the ability of an antibody of the invention to bind ERBB3 from a different species. For example, an antibody of the invention that binds human ERBB3 may also bind ERBB3 of another species. Cross-reactivity is measured by detecting specific reactivity with purified antigen in binding assays (e.g. SPR and ELISA), or binding or functional interactions with cells that physiologically express ERBB3. Methods of determining cross-reactivity include standard binding assays as described herein, such as Surface Plasmon Resonance (SPR) analysis, or flow cytometry.

The terms "inhibit" or "block" are used interchangeably and encompass both partial and complete inhibition/blocking. Inhibition/blocking of a ligand preferably reduces or alters the normal level or type of activity that occurs in the absence of inhibition or blocking when ligand binding occurs. Inhibition and blocking is also intended to include any measurable decrease in ligand binding affinity when contacted with an anti-ERBB 3 antibody compared to a ligand not contacted with an anti-ERBB 3 antibody.

The term "inhibit growth" (e.g., in reference to a cell) is intended to include any measurable reduction in cell growth.

The terms "induce an immune response" and "enhance an immune response" are used interchangeably and refer to stimulation (i.e., passive or adaptive) of an immune response to a particular antigen. The term "induction" with respect to induction of CDC or ADCC refers to stimulation of a specific direct cell killing mechanism.

The term "ADCC", i.e., antibody-dependent cell-mediated cytotoxicity, as used herein refers to the direct killing of antibody-coated target cells by Fc fragments of cells expressing Fc receptors through recognition of the antibody. The ADCC effector function of an antibody may be enhanced or reduced or eliminated by modification of the Fc-fragment of the IgG. The modification refers to mutation in the heavy chain constant region of the antibody.

The engineered antibodies or antigen binding fragments of the invention can be prepared and purified using conventional methods. The cDNA sequence of the corresponding antibody can be cloned and recombined into the GS expression vector. Recombinant immunoglobulin expression vectors can stably transfect CHO cells. As a more recommended prior art, mammalian expression systems lead to glycosylation of antibodies, particularly at the highly conserved N-terminus of the FC region. Stable clones were obtained by expression of antibodies that specifically bind to the human antigen. Positive clones were expanded in bioreactor serum-free medium to produce antibodies. The antibody-secreting culture medium can be purified and collected by conventional techniques. The antibody can be concentrated by filtration by a conventional method. Soluble mixtures and polymers can also be removed by conventional methods, such as molecular sieves, ion exchange. The resulting product is either immediately frozen, e.g., -70 ℃, or lyophilized.

"administration," "administering," and "treatment," when applied to an animal, human, experimental subject, cell, tissue, organ, or biological fluid, refer to contact of an exogenous drug, therapeutic agent, diagnostic agent, or composition with the animal, human, subject, cell, tissue, organ, or biological fluid. "administration," "administering," and "treating" may refer to, for example, therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. The treatment of the cells comprises contacting the reagent with the cells and contacting the reagent with a fluid, wherein the fluid is in contact with the cells. "administering", "administering" and "treating" also mean treating, for example, a cell in vitro and ex vivo by an agent, a diagnostic, a binding composition, or by another cell. "treatment" when applied to a human, veterinary or research subject refers to therapeutic treatment, prophylactic or preventative measures, research and diagnostic applications.

By "treating" is meant administering a therapeutic agent, such as comprising any of the antibodies of the invention, either internally or externally to a patient who has one or more symptoms of a disease for which the therapeutic agent is known to have a therapeutic effect. Typically, the therapeutic agent is administered in the subject patient or population in an amount effective to alleviate one or more symptoms of the disease, whether by inducing regression of such symptoms or inhibiting the development of such symptoms to any clinically useful degree. The amount of therapeutic agent effective to alleviate any particular disease symptom (also referred to as a "therapeutically effective amount") can vary depending on a variety of factors, such as the disease state, age, and weight of the patient, and the ability of the drug to produce a desired therapeutic effect in the patient. Whether a disease symptom has been reduced can be assessed by any clinical test commonly used by physicians or other health professional to assess the severity or progression of the symptom. Although embodiments of the invention (e.g., methods of treatment or articles of manufacture) may be ineffective in alleviating the symptoms of the disease of interest for each patient, they should alleviate the symptoms of the disease of interest in a statistically significant number of patients as determined by any statistical test known in the art, such as Student's t-test, chi-square test, U-test by Mann and Whitney, kruskal-Wallis test (H-test), jonckheere-Terpstra test, and Wilcoxon test.

The term "consisting essentially of … …" or variations thereof as used throughout the specification and claims is meant to encompass all such elements or groups of elements, and optionally includes other elements of similar or different nature than the elements, which other elements do not materially alter the basic or novel properties of a given dosing regimen, method or composition.

The term "naturally occurring" as applied to an object in accordance with the present invention refers to the fact that the object may be found in nature. For example, a polypeptide sequence or polynucleotide sequence that is present in an organism (including viruses) that can be isolated from a source in nature and which has not been intentionally modified by man in the laboratory is naturally occurring.

An "effective amount" comprises an amount sufficient to ameliorate or prevent a symptom or condition of a medical condition. An effective amount also means an amount sufficient to allow or facilitate diagnosis. The effective amount for a particular patient or veterinary subject may vary depending on the following factors: such as the condition to be treated, the general health of the patient, the method and dosage of administration, and the severity of side effects. An effective amount may be the maximum dose or dosage regimen that avoids significant side effects or toxic effects.

"exogenous" refers to a substance that is to be produced outside an organism, cell, or human body by context.

"endogenous" refers to a substance produced in a cell, organism, or human body by background.

"homology" refers to sequence similarity between two polynucleotide sequences or between two polypeptides. When a position in both of the two compared sequences is occupied by the same base or amino acid monomer subunit, e.g., if each position of two DNA molecules is occupied by adenine, then the molecules are homologous at that position. The percent homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the number of positions compared x 100%. For example, two sequences are 60% homologous if there are 6 matches or homologies at 10 positions in the two sequences when the sequences are optimally aligned. In general, comparisons are made when aligning two sequences to obtain the greatest percentage of homology.

As used herein, the expressions "cell," "cell line," and "cell culture" are used interchangeably, and all such designations include progeny thereof. Thus, the words "transformant" and "transformed cell" include the primary test cell and cultures derived therefrom, regardless of the number of transfers. It is also understood that all progeny may not be precisely identical in DNA content due to deliberate or inadvertent mutations. Mutant progeny that have the same function or biological activity as screened for in the originally transformed cell are included. Where different names are intended, they are clearly visible from the context.

"optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs or does not. For example, "optionally comprising 1-3 antibody heavy chain variable regions" means that antibody heavy chain variable regions of a particular sequence may, but need not, be present.

"pharmaceutical composition" means a composition containing one or more antibodies or antigen-binding fragments thereof described herein, as well as other components such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate administration to an organism, facilitate absorption of the active ingredient and exert biological activity.

2. Examples of the embodiments

The present invention will be further described with reference to the following examples, which are not intended to limit the scope of the present invention. The experimental methods in the examples of the present invention, in which specific conditions are not specified, are generally performed under conventional conditions such as the antibody technique laboratory manual of cold spring harbor, molecular cloning manual; or according to the conditions recommended by the manufacturer of the raw material or the goods. Reagents of specific sources are not indicated, and conventional reagents are purchased in the market.

Example 1: screening and construction of fully human antibody

ERBB3 antigen design:

the amino acid sequence of human ERBB3 protein shown in SEQ ID NO. 14 is used as the template for the antigen design. The following ERBB3 antigens not specifically mentioned refer to human ERBB3.

ERBB3 full-length protein: the sequence of ERBB3 (SEQ ID NO: 14) is as follows:

ERBB3 antigen used for screening was a commercial product, biotinylated human ERBB3-His tag (Nano biological, cat # 10201-H08H-B); the ERBB3 antigen used for detection was human ERBB3-His tag (Sino biological, cat # 10201-H08H), which is a commercial product. The detection ERBB3 antigen is the ECD region sequence of human ERBB3, the C end is provided with a His label, and the screening ERBB3 antigen is biotinylated on the basis. The ECD sequence of human ERBB3 is as follows:

the monkey ERBB3 antigen (SEQ ID NO: 16) used for detection is a commercial product Rhesuus ERBB3 Protein-His Tag (Sino biological, cat # 90043-K08H), and the sequence of the ECD region of the monkey ERBB3 antigen is added with a His Tag at the C end and has the following sequence:

collecting PBMC of multiple persons, separating B cells, extracting RNA, reverse transcribing to cDNA, and constructing natural phage surface Fab display library with cDNA as template (library capacity 1.6X 10) ¹¹ ). And packaging the constructed natural Fab phage library to form phage particles, performing panning by a liquid phase method, combining the phage with biotinylated ERBB3 liquid phase, and separating by streptavidin magnetic beads. To obtain positive sequences that bind to human ERBB3, 2-3 rounds of panning were performed with biotinylated human ERBB3, 384 monoclonal colonies were picked and packaged into phage display Fab fragments for ELISA testing. Testing monoclonal phages for binding activity to human ERBB 3: respectively coating 1 mu g/mL ERBB3 on an ELISA plate, adding phage supernatant, and finally detecting by anti-human IgG Fab HRP; OD measured by ELISA ₄₅₀ Sequencing 86 positive clones with the value greater than 0.2, analyzing the sequence to obtain the unique sequences: 45 VH (heavy chain variable region) and 51 VL (light chain variable region). Respectively connecting the obtained heavy chain and light chain variable region sequences with human IgG1 heavy chain constant region and light chain constant region sequences to obtain full-length antibodies, and performing experimental evaluation on the full-length antibodies to finally determine 1 antibody Ab1 with better binding force and function; wherein, the sequences of the connected human IgG1 heavy chain constant region and the light chain constant region are respectively shown as SEQ ID NO. 17 and SEQ ID NO. 18.

To further improve the affinity and stability of the Ab1 antibody, the antibody was subjected to S57E (Kabat numbering) site-directed mutagenesis of the HCDR2 of the antibody to form a new HCDR2 (SEQ ID NO: 3) according to the calculation model prediction, and thus antibody Ab2 was obtained.

TABLE 1 CDR sequences of the heavy and light chain variable regions of an antibody

TABLE 2 heavy and light chain variable region sequences of antibodies

Note: the CDR regions are underlined.

TABLE 3 heavy and light chain sequences of antibodies

TABLE 4 sequence numbering of antibodies and their heavy, light, variable regions

Antibody numbering	HCVR	HC	LCVR	LC
Ab1	SEQ ID NO：8	SEQ ID NO：11	SEQ ID NO：10	SEQ ID NO：13
Ab2	SEQ ID NO：9	SEQ ID NO：12	SEQ ID NO：10	SEQ ID NO：13

The IgG1 heavy and light chain constant region sequences are as follows:

example 2: preparation of fully human antibody

cDNA fragments were synthesized based on the amino acid sequences of the Ab1 and Ab2 antibody light and heavy chains and inserted into pcDNA3.1 expression vector (Life Technologies Cat. No. V790-20). Expression vectors and transfection reagent PEI (Polysciences, inc. Cat. No. 23966) were transfected into HEK293 cells (Life Technologies Cat. No. 11625019) at a ratio of 1:2 and placed in CO ₂ Incubating in incubator for 4-5 days. Collecting cell culture solution, centrifuging, filtering, loading to an antibody purification affinity column, washing the column with phosphate buffer, eluting with glycine-HCl buffer (pH2.7.0.1M Gly-HCl), neutralizing with 1M Tris-HCl pH 9.0, and dialyzing with phosphate buffer to obtain the antibody protein of the invention, wherein the concentration and purity of the antibody protein are shown in Table 5:

TABLE 5 concentration and purity of the antibody

Antibody numbering	Concentration (mg/mL)	Purity (%)
Ab1	0.5	97％
Ab2	0.4	95％

Example 3: in vitro binding Activity of fully human antibodies

3.1 in vitro indirect ELISA binding assay:

human ERBB3His protein (Sino biological, cat # 10201-H08H), monkey ERBB3His protein (Sino biological, cat # 90043-K08H), murine ERBB3His protein (Sino biological, cat # 51003-M08H), human EGFR His protein (Sino biological, cat # 10001-H08H), human ERBB2 His protein (Sino biological, cat # 10004-H08H), human ERBB4 His protein (Sino biological, cat # 10363-H08H) were each diluted to a concentration of 0.5. Mu.g/mL with PBS at pH7.4, added to a 96-well high affinity microplate at a volume of 100. Mu.L/well and incubated overnight at 4 ℃ in a refrigerator (16-20 hours). After washing the plate 3 times with PBST (0.05% Tween-20 in PBS, pH 7.4), 200. Mu.L/well of 1% Bovine Serum Albumin (BSA) blocking solution diluted with PBST was added, and the plate was incubated at 37 ℃ for 2 hours for blocking. After blocking was complete, the blocking solution was discarded and the plate was washed 1 time with PBST buffer.

The test antibody was diluted with 1% BSA in PBST, starting at 10nM, diluted with a 3-fold gradient, 11 doses, incubated at 37 ℃ for 1 hour in 100. Mu.L/Kong Jiadao enzyme plates. After completion of incubation, the plate was washed 3 times with PBST, 200. Mu.L/well of a secondary antibody anti-human HRP (Abcam, cat # ab 97225) diluted with 1% BSA in PBST was added, and incubated at 37 ℃ for 0.5 hours. After washing the plate 5 times with PBST, 100. Mu.L/well of TMB chromogenic substrate (cat # S0025, suzhou subfamily chemical reagents GmbH) was added, incubated at 25 ℃ for 8 to 15 minutes in the dark, stopped by adding 50. Mu.L/well of 1M HCl, and analyzed by reading the absorbance at 450nm with a microplate reader (Thermo, lux).

As a result of concentration signal value curve analysis, ab1 antibody showed good affinity for human-derived and monkey-derived ERBB3, and did not bind to murine ERBB3 and other ERBB receivers (EGFR, ERBB2, ERBB 3), as shown in the following table.

TABLE 6 affinity of antibodies to antigenic proteins

Note: "+" means bonded, "-" means not bonded

3.2 in vitro cell binding assay:

HEK293T cells overexpressing human ERBB3 (293T-human ERBB 3), trypsinized, harvested by centrifugation, adjusted for cell density using FACS buffer (1 XPBS containing 2 FBS), and plated onto 96-well U-bottom plates at 1X 10/well ⁵ To 2X 10 ⁵ And (4) cells. Centrifuging: 1200g, 5 min, discard the supernatant, add 100. Mu.L of antibody solution diluted with a FACS buffer gradient and incubate for 1 h at 4 ℃. Centrifuging: 1200g, 5 min, discard the supernatant, wash the cells 2 times with FACS buffer, add the fluorescence labeled secondary Antibody PE anti-human IgG Fc Antibody (Biolegged, cat # 409304) formulated in FACS buffer, resuspend the cells at 100. Mu.L per well, incubate for 1 h at 4 ℃. Centrifuging: 1200g, 5 min, discard the supernatant. After FACS buffer washing cells 2 times, re-suspended in PBS, fluorescent signal was detected using flow cytometer Bio-Rad (ZE 5), and EC of antibody-bound cells was analyzed as a curve ₅₀ And (4) concentration.

TABLE 7 binding of antibodies to 293T-hERBB3 cells (EC) ₅₀ Value)

The results show that: the antibodies of the invention can specifically bind to cells expressing human ERBB3.

Example 4: endocytosis of fully human antibodies

Whether the antibody can be endocytosed into cells together with human ERBB3 after being combined with ERBB3 is detected, and human mammary gland adenocarcinoma cells SK-Br-3 expressing ERBB3 are used for evaluation. Cells were trypsinized, harvested and resuspended in a precooled FACS buffer to a cell concentration of 2X 10 ⁶ and/mL. Taking an EP tube, adding 1mL of cell suspension, centrifuging at 1500rpm for 5 minutes, then discarding the supernatant, adding 1mL of prepared antibody to be detected for resuspension of cells, wherein the final concentration of the antibody is 20 mu g/mL, incubating for 1 hour at 4 ℃ in a shaking table, centrifuging, discarding the supernatant (4 ℃,1500rpm multiplied by 5 minutes), washing twice by a FACS buffer, and discarding the supernatant. Add 1mL of fluorescently labeled secondary Antibody working solution PE anti-human IgG Fc Antibody (Biolegend, cat # 409304) to each tube, resuspend the cells, incubate at 4 ℃ for 30 minutes in a shaker, centrifuge and discard the supernatant (4 ℃,1500 rpm. Times.5 minutes), wash twice with a FACS buffer, and discard the supernatant. Add 1mL of pre-warmed cell culture medium to each tube to resuspend the cells and mix well, divide into 4 tubes, 250 μ L each tube, 0 min group, blank group, 15 min group and 30 min group, take out 0 min and blank on ice, place the rest in 37 ℃ incubator, endocytose 15 min and 30 min respectively, take out EP tube at corresponding time point, place on ice for precooling 5 min, centrifuge all treatment groups to discard the supernatant (4 ℃,1500rpm x 5 min), wash once with FACS buffer, discard the supernatant. To all treatment groups except 0 minute group, 250. Mu.L strip buffer was added to the EP tube, incubated at room temperature for 8 minutes, centrifuged to discard the supernatant (4 ℃,1500 rpm. Times.5 minutes), washed twice with FACS buffer, and discarded the supernatant. All treatment groups were resuspended in 100. Mu.L PBS and examined by flow cytometry Bio-Rad (ZE 5).

Percent antibody internalization (%) = (fluorescence intensity value-mean fluorescence intensity value of Blank group at each time point)/(mean fluorescence lightness value of 0 minute group-mean fluorescence intensity value of Blank group) × 100, and the specific results are shown in table 8:

TABLE 8 endocytosis of fully human antibodies in tumor cells

The result shows that the human antibody human breast adenocarcinoma cell SK-Br-3 of the invention has obvious endocytosis.

Claims (21)

1. An anti-ERBB 3 antibody or antigen-binding fragment thereof, comprising: antibody heavy chain variable region and light chain variable region; wherein the variable region of the antibody heavy chain comprises at least 1 HCDR selected from the group consisting of SEQ ID NOs: 1,2, 3 or 4; the variable region of the antibody light chain comprises at least 1 LCDR selected from the group consisting of SEQ ID NOs: 5, 6 or 7.
2. The anti-ERBB 3 antibody or antigen-binding fragment thereof of claim 1, wherein,

   the heavy chain variable region of the antibody comprises HCDR1 shown in SEQ ID NO.1, HCDR2 shown in SEQ ID NO.2 and HCDR3 shown in SEQ ID NO. 4; or the like, or, alternatively,

   the heavy chain variable region of the antibody comprises HCDR1 shown in SEQ ID NO.1, HCDR2 shown in SEQ ID NO. 3 and HCDR3 shown in SEQ ID NO. 4.
3. The anti-ERBB 3 antibody or antigen-binding fragment thereof of claim 1, wherein the antibody light chain variable region comprises LCDR1 shown in SEQ ID NO. 5, LCDR2 shown in SEQ ID NO. 6, and LCDR3 shown in SEQ ID NO. 7.
4. The anti-ERBB 3 antibody or the antigen-binding fragment thereof of claim 1, which comprises an antibody heavy chain variable region and an antibody light chain variable region, wherein the antibody heavy chain variable region comprises HCDR1 of SEQ ID NO:1, HCDR2 of SEQ ID NO:2, and HCDR3 of SEQ ID NO:4; the variable region of the antibody light chain comprises LCDR1 shown in SEQ ID NO. 5, LCDR2 shown in SEQ ID NO. 6 and LCDR3 shown in SEQ ID NO. 7; or the like, or, alternatively,

   the variable region of the antibody heavy chain comprises: HCDR1 shown in SEQ ID NO.1, HCDR2 shown in SEQ ID NO. 3, and HCDR3 shown in SEQ ID NO. 4; the antibody light chain variable region comprises: LCDR1 shown in SEQ ID NO. 5, LCDR2 shown in SEQ ID NO. 6, and LCDR3 shown in SEQ ID NO. 7.
5. The anti-ERBB 3 antibody or antigen-binding fragment thereof of claim 1, further comprising a heavy chain constant region comprising a heavy chain derived from human IgG1, igG2, igG3, or IgG4, or a mutant thereof;

   preferably, the anti-ERBB 3 antibody or antigen-binding fragment thereof further comprises a heavy chain constant region derived from human IgG1 or a variant thereof;

   more preferably comprises a human IgG1 heavy chain constant region;

   most preferably, the heavy chain constant region as set forth in SEQ ID NO 17;

   optionally, the anti-ERBB 3 antibody or antigen-binding fragment thereof further comprises a light chain constant region derived from a human kappa chain, lambda chain, or mutant thereof;

   preferably a light chain constant region derived from a human lambda chain;

   most preferably, it comprises the light chain constant region as set forth in SEQ ID NO 18.
6. The anti-ERBB 3 antibody or antigen-binding fragment thereof of any one of claims 1 to 5, wherein the anti-ERBB 3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region selected from the group consisting of SEQ ID NOs: 8 or 9, or a heavy chain variable region having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% homology thereto.
7. The anti-ERBB 3 antibody or antigen-binding fragment thereof of any one of claims 1 to 6, wherein the ERBB3 antibody or antigen-binding fragment thereof comprises a light chain variable region of SEQ ID No. 10, or a light chain variable region having at least 70%, 75%, 80%, 85%, 90%, 95% or 99% homology thereto.
8. The anti-ERBB 3 antibody or antigen-binding fragment thereof of any one of claims 1 to 7, wherein the anti-ERBB 3 antibody or antigen-binding fragment thereof comprises a heavy chain selected from the group consisting of: SEQ ID NO: 11. 12, or a full-length heavy chain having at least 80%, 85%, 90%, 95%, or 99% homology thereto.
9. The anti-ERBB 3 antibody or antigen-binding fragment thereof of any one of claims 1 to 8, wherein the anti-ERBB 3 antibody or antigen-binding fragment thereof comprises a light chain of SEQ ID NO:13, or a full length light chain having at least 80%, 85%, 90%, 95%, or 99% homology thereto.
10. The anti-ERBB 3 antibody or antigen-binding fragment thereof of any of claims 1 to 9, wherein the anti-ERBB 3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region of SEQ ID No. 8, and a light chain variable region of SEQ ID No. 10; or the like, or, alternatively,

    the anti-ERBB 3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region shown in SEQ ID NO. 9 and a light chain variable region shown in SEQ ID NO. 10.
11. The anti-ERBB 3 antibody or antigen-binding fragment thereof of claim 10, wherein,

    the anti-ERBB 3 antibody comprises a heavy chain shown as SEQ ID NO.11 and a light chain shown as SEQ ID NO. 13; or the like, or, alternatively,

    the anti-ERBB 3 antibody comprises a heavy chain shown as SEQ ID NO. 12 and a light chain shown as SEQ ID NO. 13.
12. A polynucleotide encoding the anti-ERBB 3 antibody or antigen-binding fragment thereof of any one of claims 1-11.
13. An expression vector comprising the polynucleotide of claim 12.
14. A host cell into which the expression vector of claim 13 is introduced or which contains the expression vector.
15. The host cell of claim 14, wherein said host cell is a bacterial, yeast or mammalian cell; preferably E.coli, pichia, CHO cells or HEK293 cells.
16. A method of producing an anti-ERBB 3 antibody or antigen-binding fragment thereof, comprising the steps of:

    a) Culturing the host cell of any one of claims 14-15;

    b) Isolating the antibody from the culture; and the number of the first and second groups,

    c) Purifying the antibody.
17. A pharmaceutical composition comprising the anti-ERBB 3 antibody, or antigen-binding fragment thereof, of any one of claims 1-11, and a pharmaceutically acceptable excipient, diluent, or carrier.
18. A detection or diagnostic reagent comprising the anti-ERBB 3 antibody, or antigen-binding fragment thereof, of any one of claims 1-11, and an excipient, diluent, or carrier useful for detection or diagnosis.
19. Use of an anti-ERBB 3 antibody or antigen-binding fragment thereof according to any one of claims 1-11, or a composition according to claim 17, in the manufacture of a medicament for the treatment or prevention of an ERBB3 mediated disease or disorder.
20. Use of an anti-ERBB 3 antibody or antigen-binding fragment thereof according to any one of claims 1-11, or a detection or diagnostic reagent according to claim 18, in the preparation of a kit for the detection, diagnosis, prognosis of an ERBB 3-mediated disease or disorder.
21. The use of claims 19-20, wherein:

    the disease or disorder is cancer;

    preferably a cancer in which ERBB3 is expressed or overexpressed;

    more preferably breast cancer, ovarian cancer, prostate cancer, endometrial cancer, thyroid cancer, renal cancer, lung cancer, gastric cancer, colon cancer, bladder cancer, cervical cancer, gallbladder cancer, pancreatic cancer, testicular cancer, soft tissue sarcoma, head and neck cancer, glioma or melanoma.