Patent application title: DE-IMMUNISED ANTI-ERBB3 ANTIBODIES
Inventors:
IPC8 Class: AC07K1632FI
USPC Class:
1 1
Class name:
Publication date: 2021-06-24
Patent application number: 20210189003
Abstract:
Provided herein are antibody molecules that bind specifically to ERBB3
and related nucleic acid molecules, vectors and host cells. Also provided
herein are medical uses of such antibody molecules.Claims:
1. A nucleic acid molecule encoding the heavy chain variable (VH) region
amino acid sequence and the light chain variable (VL) region amino acid
sequence of an anti-ERBB3 antibody or an antigen-binding portion thereof,
wherein (a) the VH region amino acid sequence comprises HCDR1 of
GFTFSDYSMS (SEQ ID NO: 24), HCDR2 of VSTISDSGTYTYYPDSVKG (SEQ ID NO: 25)
and HCDR3 of EWGDYDGFDF (SEQ ID NO: 15); and the VL region amino acid
sequence comprises LCDR1 of RASQEISTYLS (SEQ ID NO: 261), LCDR2 of
AASTLQS (SEQ ID NO:26) and LCDR3 of LQYDSSPLT (SEQ ID NO: 262); or (b)
the VH region amino acid sequence comprises HCDR1 of GFTFSDYSMS (SEQ ID
NO: 24), HCDR2 of VSTISDSGTYTYYPDSVKG (SEQ ID NO: 25) and HCDR3 of
EWGDYDGFDF (SEQ ID NO: 15); and the VL region amino acid sequence
comprises LCDR1 of RASQEISSYLS (SEQ ID NO: 21), LCDR2 of AASSLDT (SEQ ID
NO: 263) and LCDR3 of amino acids 89-97 of SEQ ID NO: 224.
2. The nucleic acid molecule of claim 1, wherein (a) the VH region amino acid sequence comprises SEQ ID NO:236 and the VL region amino acid sequence comprises SEQ ID NO:225; or (b) the VH region amino acid sequence comprises SEQ ID NO:235 and the VL region amino acid sequence comprises SEQ ID NO:224.
3. The nucleic acid molecule of claim 1, wherein the antibody or antigen-binding portion is humanized or chimeric.
4. The nucleic acid molecule of claim 1, wherein the VH region, the VL region, or both the VH and the VL region comprise one or more human framework region amino acid sequences.
5. The nucleic acid molecule of claim 1, wherein the VH region, the VL region, or both the VH and the VL region comprise a human variable region framework scaffold amino acid sequence into which the CDRs have been inserted.
6. The nucleic acid molecule of claim 1, wherein the VH region comprises an IGHV3-11 human germline scaffold amino acid sequence into which the HCDR1, HCDR2 and HCDR3 amino acid sequences have been inserted.
7. The nucleic acid molecule of claim 1, wherein the VL region comprises an IGKV1-39 human germline scaffold amino acid sequence into which the LCDR1, LCDR2 and LCDR3 amino acid sequences have been inserted.
8. The nucleic acid molecule of claim 1, wherein the nucleic acid molecule further encodes an immunoglobulin constant region.
9. The nucleic acid molecule of claim 8, wherein the immunoglobulin constant region is IgG, IgE, IgM, IgD or IgA.
10. The nucleic acid molecule of claim 9, wherein the immunoglobulin constant region is IgG1, IgG2, IgG3, IgG4, IgA1 or IgA2.
11. The nucleic acid molecule of claim 8, wherein the immunoglobulin constant region is immunologically inert.
12. The nucleic acid molecule of claim 8, wherein the immunoglobulin constant region is a wild-type human IgG4 constant region, a human IgG4 constant region comprising the amino acid substitution S228P, a wild-type human IgG1 constant region, a human IgG1 constant region comprising the amino acid substitutions L234A, L235A and G237A or a wild-type human IgG2 constant region, wherein numbering is according to the EU index as in Kabat.
13. The nucleic acid molecule of claim 8, wherein the immunoglobulin constant region comprises any one of SEQ ID NOs: 239-245.
14. The nucleic acid molecule of claim 1, wherein the antibody or antigen-binding portion is an Fab, an Fab', an F(ab')2, an Fv, an scFv, a diabody, a triabody, a tetrabody or a bis-scFv.
15. An expression vector comprising the nucleic acid molecule of claim 1.
16. A recombinant host cell comprising the nucleic acid molecule of claim 1.
17. A recombinant host cell comprising the expression vector of claim 15.
Description:
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a division of U.S. patent application Ser. No. 16/925,570, filed on Jul. 10, 2020, which is a continuation of International Patent Application No. PCT/EP2019/056506, filed on Mar. 14, 2019, which claims the benefit of GB Patent Application No. 1804094.9, filed on Mar. 14, 2018, the disclosure of each of which is hereby incorporated by reference in its entirety.
DESCRIPTION OF THE TEXT FILE SUBMITTED ELECTRONICALLY
[0002] The contents of the text file submitted electronically herewith are incorporated herein by reference in their entirety: A computer readable format copy of the Sequence Listing (filename: ULTH_001_02US_SeqList_ST25.txt, date recorded: Mar. 1, 2021, file size .about.141,322 bytes).
FIELD OF THE INVENTION
[0003] The invention relates to antibody molecules binding specifically to ERBB3 (also known as ErbB-3, HER3, LCCS2, MDA-BF-1, c-erbB-3, c-erbB3, erbB3-S, p180-ErbB3, p45-sErbB3, p85-sErbB3, erb-b2 receptor tyrosine kinase 3) and medical uses thereof.
BACKGROUND OF THE INVENTION
[0004] ERBB3 (also known as ErbB-3, HER3, LCCS2, MDA-BF-1, c-erbB-3, c-erbB3, erbB3-S, p180-ErbB3, p45-sErbB3, p85-sErbB3, and erb-b2 receptor tyrosine kinase 3) is a transmembrane receptor tyrosine kinase (RTK) protein that belongs to the immunoglobulin superfamily. ERBB3 lacks significant tyrosine kinase activity of its own, and is activated via heterodimerization with other related RTKs such as HER2, EGFR, and MET. This heterodimerization is primarily driven by the predominant ligand for ERBB3, which is Heregulin (HRG), also known as Neuregulin 1 (NRG1). ERBB3-HRG interaction triggers the tyrosine phosphorylation of ERBB3 by heterodimer partners and the activation of diverse intracellular signaling networks. Importantly, ERBB3 is the most potent activator of the PI3K/AKT signaling pathway in the EGFR family.
[0005] The ERBB3 receptor is often overexpressed in tumours of the head and neck, lung, breast, ovary, prostate, colon, pancreas, and gastrointestinal tract. The overexpression of ERBB3 is strongly linked with poor prognosis, and ERBB3 is believed to be influential in resistance mechanisms to radiotherapy and various chemotherapeutic and biotherapeutic drugs. As the preferred dimerization partner of HER2, amplified ERBB3 signaling in HER2+ breast tumours is believed to be partly responsible for resistance to trastuzumab therapy. Therapeutic antibodies that antagonise ERBB3 signaling by blocking its ability to dimerise with other key receptors have the potential to mediate anti-tumour effects, via two mechanisms: 1. Potent inhibition of the ERBB3 signaling pathway by locking the receptors into a monomeric form. 2. Antibody effector-function mediated engagement of immune cells.
[0006] The majority of currently approved antibody therapeutics are derived from immunized rodents. Many of those antibodies have undergone a process known as "humanization", via the "grafting" of murine Complementarity-Determining Regions (CDRs) into human v-gene framework sequences (see Nelson et al., 2010, Nat Rev Drug Discov 9: 767-774). This process is often inaccurate and leads to a reduction in target binding affinity of the resulting antibody. To return the binding affinity of the original antibody, murine residues are usually introduced at key positions in the variable domain frameworks of the grafted v-domains (also known as "back-mutations").
[0007] While antibodies humanized via CDR grafting and back mutations have been shown to induce lower immune response rates in the clinic in comparison to those with fully murine v-domains, antibodies humanized using this basic grafting method still carry significant clinical development risks due to the potential physical instability and immunogenicity motifs still housed in the grafted CDR loops. Antibodies such as anti-ERBB3, which potentially engage immune effector functions as part of their mechanism of action, are at particularly high risk of immunogenicity as they can encourage phagocytosis of ERBB3+ target cells, leading to antigen processing of the antibody along with the target cell. As animal testing of protein immunogenicity is often non-predictive of immune responses in man, antibody engineering for therapeutic use focuses on minimizing predicted human T-cell epitope content, non-human germline amino acid content and aggregation potential in the purified protein.
[0008] The ideal humanized antagonistic anti-ERBB3 antibody would therefore have as many residues as possible in the v-domains that are identical to those found in both the frameworks and CDRs of well-characterized human germline sequences. This high level of identity to high-stability germlines that are highly expressed in the maximum number of potential patients minimises the risk of a therapeutic antibody having unwanted immunogenicity in the clinic, or unusually high `cost of goods` in manufacturing.
[0009] Townsend et al. (2015; PNAS 112: 15354-15359) describe a method for generating antibodies in which CDRs derived from rat, rabbit and mouse antibodies were grafted into preferred human frameworks and then subject to a human germ-lining approach termed "Augmented Binary Substitution". Although the approach demonstrated a fundamental plasticity in the original antibody paratopes, in the absence of highly accurate antibody-antigen co-crystal structural data, it is still not possible to reliably predict which individual residues in the CDR loops of any given antibody can be converted to human germline, and in what combination. Additionally, the Townsend et al. study did not address the addition of mutagenesis beyond the residues found in the human germline at positions where the removal of development risk motifs might be beneficial. This is a technological limitation which renders the process inherently inefficient, requiring an extra stage of modification of the starting antibody sequence. In addition, it cannot currently be accurately predicted what modifications in distal positions of the protein sequence of an individual v-domain, or even on the partner v-domain, might facilitate the removal of risk motifs while maintaining antigen binding affinity and specificity.
[0010] CDR germ-lining and development quality optimisation is thus a complex, multifactorial problem, as multiple functional properties of the molecule should preferably be maintained, including in this instance: target binding specificity, affinity to ERBB3 from both human and animal test species (e.g. rhesus monkey, also known as the rhesus macaque, i.e. Macaca mulatta), v-domain biophysical stability and/or IgG yield from protein expression platforms used in research, clinical and commercial supply. Antibody engineering studies have shown that mutation of even single residue positions in key CDRs can have dramatic effects on all of these desired molecular properties.
[0011] WO2011136911A2 describes an antagonistic murine anti-ERBB3 IgG molecule termed "24C05", and also the preparation of humanized forms (h24C05). Those humanized forms of 24C05 were produced using classical humanization techniques, i.e. by grafting of Kabat-defined murine CDRs into human heavy and light chain framework sequences, with some of the human framework residues being potentially back-mutated to the correspondingly positioned 24C05 murine residues. For reasons noted above, such humanized forms of 24C05 described in WO2011136911A2 are not ideal.
SUMMARY OF THE INVENTION
[0012] The present invention provides a number of anti-ERBB3 antibodies and medical uses thereof.
[0013] According to one aspect of the invention, there is provided an antibody molecule which specifically binds to human ERBB3, and optionally also to rhesus monkey ERBB3, or an antigen-binding portion thereof, wherein the antibody molecule or antigen-binding portion comprises a heavy chain variable region with:
[0014] an HCDR1 having amino acids in sequence in the following order: G-F-T-F-S-D-Y-G or any amino acid (such as S)-M-S (SEQ ID NO:1);
[0015] an HCDR2 having amino acids in sequence in the following order: V-S-T-I-S-D-G or any amino acid (such as S, D)-G-T or a conservative substitution of T (such as S)-Y or any amino acid (such as T)-T or any amino acid (such as I)-Y-Y-P or any amino acid (such as A)-D-N or a conservative substitution of N (such as S)-V-K-G (SEQ ID NO:2); and
[0016] an HCDR3 having amino acids in sequence in the following order: E or any amino acid (such as M)-W or any amino acid (such as F, L, M, Q or Y)-G-D-Y or any amino acid (such as A, D, E, H, L, M, N, Q, S, T or VV)-D-G-F or any amino acid (such as I, L, W, Y)-D-Y or any amino acid (such as A, D, E, F, H, I, K, L, M, N, Q, R, S, V, W) (SEQ ID NO:3).
[0017] In aspects of the invention, the HCDR1 of the antibody molecule or antigen-binding portion may exclude the sequence GFTFSDYAMS (SEQ ID NO:4; 24C05 murine/humanized antibody HCDR1 disclosed in WO2011136911A2; US20110256154A1), the HCDR2 of the antibody molecule or antigen-binding portion may exclude the sequence VSTISDGGTYTYYPDNVKG (SEQ ID NO:5; 24C05 murine/humanized antibody HCDR2 disclosed in WO2011136911A2; US20110256154A1), and/or the HCDR3 of the antibody molecule or antigen-binding portion may exclude the sequence EWGDYDGFDY (SEQ ID NO:6; 24C05 murine/humanized antibody HCDR3 disclosed in WO2011136911A2; US20110256154A1).
[0018] The antibody molecule or antigen-binding portion may further comprise a light chain variable region with:
[0019] an LCDR1 having amino acids in sequence in the following order: R-A-S-Q-E or any amino acid (such as S, I, N)-I-S-G or a conservative substitution of G (such as S, T)-Y-L-S or a conservative substitution of S (such as N) (SEQ ID NO:7);
[0020] an LCDR2 having amino acids in sequence in the following order: A or any amino acid (such as E)-A-S-T or a conservative substitution of T (such as S, N)-L-D or any amino acid (such as H, K, Q)-S or T (SEQ ID NO:8); and
[0021] an LCDR3 having amino acids in sequence in the following order: L or any amino acid (such as Q)-Q-Y or any amino acid (such as S)-D or any amino acid (such as Y)-S-Y or any amino acid (such as T, S)-P or any amino acid (such as H)-Y or any amino acid (such as L)-T (SEQ ID NO:9).
[0022] In aspects of the invention, the LCDR1 of the antibody molecule or antigen-binding portion may exclude the sequence RASQEISGYLS (SEQ ID NO:10; 24C05 murine/humanized antibody LCDR1 disclosed in WO2011136911A2; US20110256154A1), and/or the LCDR2 of the antibody molecule or antigen-binding portion may exclude the sequence AASTLDS (SEQ ID NO:11; 24C05 murine/humanized antibody LCDR2 disclosed in WO2011136911A2; US20110256154A1), and/or the LCDR3 of the antibody molecule or antigen-binding portion may exclude the sequence LQYDSYPYT (SEQ ID NO:12; 24C05 murine/humanized antibody LCDR3 disclosed in WO2011136911A2; US20110256154A1).
[0023] In some aspects, disclosed herein is an anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody comprises a heavy chain variable (VH) region and a light chain variable (VL) region, wherein
[0024] (a) the HCDR1 comprises the amino acid sequence G-F-T-F-S-D-Y-X.sub.1-M-S, wherein X.sub.1 is G or any other amino acid (SEQ ID NO:1);
[0025] (b) the HCDR2 comprises V-S-T-I-S-D-X.sub.1-G-X.sub.2-X.sub.3-X.sub.4-Y-Y-X.sub.5-D-X.sub.6-V-K-G- , wherein X.sub.1 is G or any other amino acid, X.sub.2 is T or a conservative substitution of T, X.sub.3 is Y or any other amino acid, X.sub.4 is T or any other amino acid, X.sub.5 is P or any other amino acid, and X.sub.6 is N or a conservative substitution of N (SEQ ID NO:2);
[0026] (c) the HCDR3 comprises X.sub.1-X.sub.2-G-D-X.sub.3-D-G-X.sub.4-D-X.sub.5, wherein X.sub.1 is E or any other amino acid, X.sub.2 is W or any other amino acid, X.sub.3 is Y or any other amino acid, X.sub.4 is F or any other amino acid, and X.sub.5 is Y or any other amino acid (SEQ ID NO:3);
[0027] (d) the LCDR1 comprises R-A-S-Q-X.sub.1-I-S-X.sub.2-Y-L-X.sub.3, wherein X.sub.1 is E or any other amino acid, X.sub.2 is G or a conservative substitution of G, and X.sub.3 is S or a conservative substitution of S (SEQ ID NO:7);
[0028] (e) the LCDR2 comprises X.sub.1-A-S-X.sub.2-L-X.sub.3-S, wherein X.sub.1 is A or any other amino acid, X.sub.2 is T or a conservative substitution of T, and X.sub.3 is or D any other amino acid (SEQ ID NO:8); and
[0029] (f) the LCDR3 comprises X.sub.1-Q-X.sub.2-X.sub.3-S-X.sub.4-X.sub.5-X.sub.6-T, wherein X.sub.1 is L or any other amino acid, X.sub.2 is Y or any other amino acid, X.sub.3 is D or any other amino acid, X.sub.4 is Y or any other amino acid, X.sub.5 is P or any other amino acid, and X.sub.6 is Y or any other amino acid (SEQ ID NO:9). In some aspects, the LCDR2 comprises X.sub.1-A-S-X.sub.2-L-X.sub.3-S (SEQ ID NO:8), wherein the seventh residue in the sequence is a conservative substitution of S (for example, T).
[0030] In some aspects, the invention provides an anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody comprises a heavy chain variable (VH) region and a light chain variable (VL) region, wherein
[0031] (a) the VH region amino acid sequence comprises HCDR1 of GFTFSDYSMS (SEQ ID NO: 24), HCDR2 of VSTISDSGTYTYYPDSVKG (SEQ ID NO: 25) and HCDR3 of EWGDYDGFDF (SEQ ID NO: 15); and the VL region amino acid sequence comprises LCDR1 of RASQEISTYLS (SEQ ID NO: 261), LCDR2 of AASTLQS (SEQ ID NO:26) and LCDR3 of LQYDSSPLT (SEQ ID NO: 262);
[0032] (b) the VH region amino acid sequence comprises HCDR1 of GFTFSDYSMS (SEQ ID NO: 24), HCDR2 of VSTISDSGTYTYYPDSVKG (SEQ ID NO: 25) and HCDR3 of EWGDYDGFDF (SEQ ID NO: 15); and the VL region amino acid sequence comprises LCDR1 of RASQEISSYLS (SEQ ID NO: 21), LCDR2 of AASSLDT (SEQ ID NO: 263) and LCDR3 of LQYDSTPYT (SEQ ID NO: 23);
[0033] (c) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDSGSYTYYPDSVKG (SEQ ID NO:19) and HCDR3 of ELGDYDGFDY (SEQ ID NO:20); and the VL region amino acid sequence comprises LCDR1 of RASQEISSYLS (SEQ ID NO:21), LCDR2 of AASSLDS (SEQ ID NO:22) and LCDR3 of LQYDSTPYT (SEQ ID NO:23);
[0034] (d) the VH region amino acid sequence comprises HCDR1 of GFTFSDYSMS (SEQ ID NO:24), HCDR2 of VSTISDSGTYTYYPDSVKG (SEQ ID NO:25) and HCDR3 of EWGDYDGFDF (SEQ ID NO:15); and the VL region amino acid sequence comprises LCDR1 of RASQEISSYLS (SEQ ID NO:21), LCDR2 of AASTLQS (SEQ ID NO:26) and LCDR3 of LQYDSTPLT (SEQ ID NO:18);
[0035] (e) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDSGSYIYYADSVKG (SEQ ID NO:14) and HCDR3 of EWGDYDGFDF (SEQ ID NO:15); and the VL region amino acid sequence comprises LCDR1 of RASQSISSYLS (SEQ ID NO:16), LCDR2 of AASSLQS (SEQ ID NO:17) and LCDR3 of LQYDSTPLT (SEQ ID NO:18);
[0036] (f) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDSGSYIYYADSVKG (SEQ ID NO:14) and HCDR3 of EWGDYDGFDH (SEQ ID NO:27); and the VL region amino acid sequence comprises LCDR1 of RASQEISSYLS (SEQ ID NO:21), LCDR2 of AASSLQS (SEQ ID NO:17) and LCDR3 of LQYDSTPLT (SEQ ID NO:18);
[0037] (g) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDGGSYTYYADSVKG (SEQ ID NO:28) and HCDR3 of EWGDYDGFDE (SEQ ID NO:29); and the VL region amino acid sequence comprises LCDR1 of RASQSISGYLS (SEQ ID NO:30), LCDR2 of AASTLQS (SEQ ID NO:26) and LCDR3 of LQYDSTPYT (SEQ ID NO:23);
[0038] (h) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDGGSYTYYADNVKG (SEQ ID NO:31) and HCDR3 of EWGDYDGFDF (SEQ ID NO:15); and the VL region amino acid sequence comprises LCDR1 of RASQSISSYLS (SEQ ID NO:16), LCDR2 of AASTLQS (SEQ ID NO:26) and LCDR3 of LQYDSTPLT (SEQ ID NO:18); or
[0039] (i) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDSGSYIYYADSVKG (SEQ ID NO:14) and HCDR3 of EWGDYDGFDE (SEQ ID NO:29); and the VL region amino acid sequence comprises LCDR1 of RASQSISSYLS (SEQ ID NO:16), LCDR2 of AASSLQS (SEQ ID NO:17) and LCDR3 of LQYDSTPLT (SEQ ID NO:18).
[0040] In some aspects, disclosed herein is an anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody comprises a heavy chain variable (VH) region and a light chain variable (VL) region, wherein
[0041] the VH region amino acid sequence comprises:
[0042] (a) HCDR1 of SEQ ID NO: 13 or 24;
[0043] (b) HCDR2 of SEQ ID NO: 14, 19, 25, 28 or 31; and
[0044] (c) HCDR3 of SEQ ID NO: 15, 20, 27 or 29; and
[0045] the VL region amino acid sequence comprises:
[0046] (a') LCDR1 of SEQ ID NO: 16, 21, 30 or 261;
[0047] (b') LCDR2 of SEQ ID NO: 17, 22, 26 or 263; and
[0048] (c') LCDR3 of SEQ ID NO: 18, 23 or 262.
[0049] In some aspects, disclosed herein is an anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody comprises a heavy chain variable (VH) region and a light chain variable (VL) region, wherein
[0050] (a) the VH region amino acid sequence comprises SEQ ID NO:236 and the VL region amino acid sequence comprises SEQ ID NO:225;
[0051] (b) the VH region amino acid sequence comprises SEQ ID NO:232 and the VL region amino acid sequence comprises SEQ ID NO:221;
[0052] (c) the VH region amino acid sequence comprises SEQ ID NO:253 and the VL region amino acid sequence comprises SEQ ID NO:254; or
[0053] (d) the VH region amino acid sequence comprises SEQ ID NO:255 and the VL region amino acid sequence comprises SEQ ID NO:256.
[0054] Also provided according to the invention is an immunoconjugate comprising the antibody molecule or antigen-binding portion thereof as defined herein linked, fused or conjugated to a therapeutic agent.
[0055] In another aspect the invention provides a nucleic acid molecule encoding the antibody molecule or antigen-binding portion thereof as defined herein.
[0056] Further provided is a vector comprising the nucleic acid molecule of the invention.
[0057] Also provided is a host cell comprising the nucleic acid molecule or the vector of the invention as defined herein.
[0058] In a further aspect there is provided a method of producing an anti-ERBB3 antibody and/or an antigen-binding portion thereof, comprising culturing the host cell of the invention under conditions that result in expression and/or production of the antibody and/or the antigen-binding portion thereof, and isolating the antibody and/or the antigen-binding portion thereof from the host cell or culture.
[0059] In another aspect of the invention there is provided a pharmaceutical composition comprising the antibody molecule or antigen-binding portion thereof of the invention as defined herein, or the nucleic acid molecule of the invention as defined herein, or the vector of the invention as defined herein.
[0060] Further provided is a method for enhancing an immune response in a subject, comprising administering an effective amount of the antibody molecule or antigen-binding portion thereof of the invention as defined herein, or the immunoconjugate of the invention as defined herein, or the nucleic acid molecule of the invention as defined herein, or the vector of the invention as defined herein, or the pharmaceutical composition of the invention as defined herein.
[0061] In a further aspect there is provided a method for treating or preventing cancer in a subject, comprising administering an effective amount of the antibody molecule or antigen-binding portion thereof of the invention as defined herein, or the immunoconjugate of the invention as defined herein, or the nucleic acid molecule of the invention as defined herein, or the vector of the invention as defined herein, or the pharmaceutical composition of the invention as defined herein.
[0062] Further provided herein is an antibody molecule or antigen-binding portion thereof as defined herein, or the immunoconjugate as defined herein, or the nucleic acid molecule as defined herein, or the vector as defined herein, or the pharmaceutical composition as defined herein, for use as a medicament. The invention also provides an antibody molecule or antigen-binding portion thereof of the invention as defined herein, or the immunoconjugate of the invention as defined herein, or the nucleic acid molecule of the invention as defined herein, or the vector of the invention as defined herein, or the pharmaceutical composition of the invention as defined herein, for use in the treatment of cancer.
[0063] In another aspect the invention provides the antibody molecule, or antigen-binding portion thereof, or the immunoconjugate, or the nucleic acid molecule, or the vector for use, or the method of treatment of the invention as defined herein, for separate, sequential or simultaneous use in a combination with a second therapeutic agent, for example an anti-cancer agent.
[0064] In a further aspect there is provided the use of an antibody molecule or antigen-binding portion thereof of the invention as defined herein, or an immunoconjugate of the invention as defined herein, or a nucleic acid molecule of the invention as defined herein, or a vector of the invention as defined herein, or a pharmaceutical composition of the invention as defined herein, in the manufacture of a medicament for the treatment of cancer.
[0065] The invention also provides a method for treating or preventing an an autoimmune disease or an inflammatory disease in a subject, comprising administering an effective amount of the antibody molecule or antigen-binding portion thereof as defined herein, or the immunoconjugate as defined here, or the nucleic acid molecule as defined herein, or the vector as defined herein, or the pharmaceutical composition as defined herein.
[0066] The autoimmune disease or inflammatory disease may be selected in all aspects from the group consisting of: arthritis, asthma, multiple sclerosis, psoriasis, Crohn's disease, inflammatory bowel disease, lupus, Grave's disease and Hashimoto's thyroiditis, and ankylosing spondylitis.
[0067] Also provided is an antibody molecule or antigen-binding portion thereof as defined herein, or the immunoconjugate as defined herein, or the nucleic acid molecule as defined herein, or the vector as defined herein, or the pharmaceutical composition as defined herein, for use in the treatment of an autoimmune disease or an inflammatory disease.
[0068] Further provided is the use of an antibody molecule or antigen-binding portion thereof as defined herein, or an immunoconjugate as defined herein, or a nucleic acid molecule as defined herein, or a vector as defined herein, or a pharmaceutical composition as defined herein, in the manufacture of a medicament for the treatment of an autoimmune disease or an inflammatory disease.
[0069] The invention also provides a method for treating or preventing a cardiovascular disease or a fibrotic disease in a subject, comprising administering an effective amount of the antibody molecule or antigen-binding portion thereof as defined herein, or the immunoconjugate as defined here, or the nucleic acid molecule as defined herein, or the vector as defined herein, or the pharmaceutical composition as defined herein.
[0070] Also provided is an antibody molecule or antigen-binding portion thereof as defined herein, or the immunoconjugate as defined herein, or the nucleic acid molecule as defined herein, or the vector as defined herein, or the pharmaceutical composition as defined herein, for use in the treatment of a cardiovascular disease or a fibrotic disease.
[0071] Further provided is the use of an antibody molecule or antigen-binding portion thereof as defined herein, or an immunoconjugate as defined herein, or a nucleic acid molecule as defined herein, or a vector as defined herein, or a pharmaceutical composition as defined herein, in the manufacture of a medicament for the treatment of an autoimmune disease, an inflammatory disease or a fibrotic disease.
[0072] The cardiovascular disease in any aspect of the invention may for example be coronary heart disease or atherosclerosis.
[0073] The fibrotic disease in any aspect of the invention may be selected from the group consisting of myocardial infarction, angina, osteoarthritis, pulmonary fibrosis, cystic fibrosis, bronchitis and asthma.
[0074] The invention also provides a method of producing an antibody molecule which specifically binds to human ERBB3 and optionally also to rhesus monkey ERBB3, or an antigen-binding portion thereof, comprising the steps of:
[0075] (1) grafting anti-ERBB3 CDRs from a non-human source into a human v-domain framework to produce a humanized anti-ERBB3 antibody molecule or antigen-binding portion thereof;
[0076] (2) generating a phage library of clones of the humanized anti-ERBB3 antibody molecule or antigen-binding portion thereof comprising one or more mutations in the CDRs;
[0077] (3) screening the phage library for binding to human ERBB3 and optionally also to rhesus monkey ERBB3;
[0078] (4) selecting clones from the screening step (3) having binding specificity to human ERBB3 and optionally also to rhesus monkey ERBB3; and
[0079] (5) producing an antibody molecule which specifically binds to human ERBB3 and optionally also to rhesus monkey ERBB3, or an antigen-binding portion thereof from clones selected from step (4).
[0080] The method may comprise a further step of producing additional clones based on the clones selected in step (4), for example based on further exploratory mutagenesis at specific positions in the CDRs of the clones selected in step (4), to enhance humanization and/or minimise human T cell epitope content and/or improve manufacturing properties in the antibody molecule or antigen-binding portion thereof produced in step (5).
BRIEF DESCRIPTION OF THE FIGURES
[0081] FIG. 1A-FIG. 1B. Direct binding ELISA and Alphascreen competition screening of library-derived anti-ERBB3 Fabs against human and rhesus ERBB3-Fc proteins. Clones were derived from multiple phage selection branches where phage populations were selected on biotinylated human, or rhesus monkey ERBB3 proteins in each of rounds II-IV. `Hammer-Hug` rounds were also performed in separate rounds II and III. After each round of selection, library-derived clones were screened as periplasmically-expressed Fab proteins, against both human (huERBB3) and rhesus (rhERBB3) in ELISA (FIG. 1A), and in blocking the binding of 24C05 IgG in binding to huERBB3 by Alphascreen (FIG. 1B). Mean.+-.SD values in each round are represented in grey bars.
[0082] FIG. 2A-FIG. 2B. Analysis of CDR residue tolerance for mutation to germline. A plot of murine amino acid retention frequencies in the CDRs of the ELISA-positive population of 658 unique Fab clones that demonstrated human and rhesus ERBB3 cross-reactivity is shown for VL (SEQ ID NOs:32-34) (FIG. 2A) and VH (SEQ ID NOs:35-37) (FIG. 2B) domains, respectively. Only those residues targeted for human/murine residue mutagenesis are plotted, other than in the HCDR3. CDR residues noted in parentheses on the X-axes were identical to those found in the human germlines used for grafting (IGKV1-39 and IGHV3-11). Those residues in the CDRs that are not in parentheses, but whose values are set at 0, were mutated to human germline during the grafting process. In both plots the dashed line in grey at 75% represents the cut off for tolerance of murine residue replacement by human germline.
[0083] FIG. 3A-FIG. 3B. Direct titration ELISA for IgG binding to human and rhesus ERBB3 proteins. Chimeric and humanized 24C05, library-derived and designer clones in human IgG1null format were titrated (in nM) in a direct binding ELISA against human (FIG. 3A) and rhesus (FIG. 3B) ERBB3-Fc proteins. All clones other than Isotype IgG1null control demonstrated binding activity against both orthologs of ERBB3.
[0084] FIG. 4. Epitope competition analysis of IgG1null proteins in Alphascreen. Anti-ERBB3 IgG1null clones were applied in an epitope competition assay using Alphascreen technology. In this assay, library-derived and designer IgGs were analysed for their retention of the h24C05 epitope by competing for h24C05 IgG1 null binding to human ERBB3 protein, in solution. All clones analysed showed strong, concentration-dependent neutralisation of h24C05 binding to ERBB3.
[0085] FIG. 5A-FIG. 5B. Flow cytometric binding to human and rhesus ERBB3+ HEK-293 cells for library-derived and primary designer leads. Chimeric and humanized 24C05, library-derived and designer leads in IgG1null format were examined for specific binding on human (FIG. 5A) and rhesus (FIG. 5B) ERBB3-transfected HEK-293 cells. IgGs were tested at concentrations ranging from 0.008-500 nM. Concentration-dependent binding was observed against both human and rhesus cell lines for all ERBB3-specific antibodies but not isotype controls. No binding signals above background were observed against wild type HEK-293 cells.
[0086] FIG. 6A-FIG. 6B. T cell epitope peptide content in lead antibody v-domains. The v-domains of h24C05 (FIG. 6A) and 15G11 (FIG. 6B) antibodies were examined for the presence of Germline (GE), High Affinity Foreign (HAF), Low Affinity Foreign (LAF) and TCED+ T cell receptor epitopes. Both the VH and VL domains of each antibody were found to contain multiple high-risk human T cell epitopes. In 15G11, despite having germline frameworks and multiple human germline residue changes in the CDRs, the high-risk epitope content was significantly increased in comparison to h24C05, rather than the expected reduction.
[0087] FIG. 7A-FIG. 7D. In silico disruption of T cell epitope peptide content in lead antibody v-domains. The v-domains High Affinity Foreign (HAF), Low Affinity Foreign (LAF) and TCED+ T cell receptor epitopes found in 15G11 were targeted for ablation. In silico mutagenesis analyses were performed to identify non-germline amino acid changes that might maintain antibody binding function but ablate one or more 9-mer peptide epitopes. There analyses were performed for peptides found in the HCDR-1 (FIG. 7A), LCDR-1 (FIG. 7B), LCDR-2 (FIG. 7C) and LCDR-3 (FIG. 7D). Residues are numbered according to the Kabat numbering scheme, the 9-mer peptide sequence is highlighted and p1 and p9 positions indicated. Favoured epitope disrupting mutations are indicated above arrows in grey, disfavoured in black. A delta symbol next to an amino acid one-letter abbreviation means that this mutation would render the new peptide a germ line (GE) peptide. An asterisk next to an amino acid one-letter abbreviation refers to a disfavoured mutation because the use of that residue would create a new isomerisation development risk motif (DG). FIG. 7A shows SEQ ID NO:257. FIG. 7B shows SEQ ID NO:258. FIG. 7C shows SEQ ID NO:259. FIG. 7D shows SEQ ID NO:260.
[0088] FIG. 8A-FIG. 8B. Direct titration ELISA for 15G11-DI IgGs binding to human and rhesus ErbB3 proteins. Chimeric and humanized 24C05, isotype control IgG1 and 15G11-DI1 to DI11 clones in human IgG1 format were titrated (in nM) in a direct binding ELISA against human (FIG. 8A) and rhesus (FIG. 8B) ERBB3-Fc proteins. All clones other than Isotype IgG1 control demonstrated binding activity against both orthologs of ERBB3.
[0089] FIG. 9. Epitope competition analysis of IgG1 proteins in Alphascreen. Chimeric and humanized 24C05, isotype control IgG1 and 15G11-DI1 to DI11 clones in human IgG1 format were applied in an epitope competition assay using Alphascreen technology. In this assay, IgGs were analysed for their retention of the same functional epitope as h24C05 by competing for h24C05 IgG1 binding to human ERBB3 protein, in solution. All clones analysed showed strong, concentration-dependent neutralisation of h24C05 binding to ERBB3, with the exception of 15G11-DI11.
[0090] FIG. 10A-FIG. 10B. Flow cytometric binding to human and rhesus ERBB3+ HEK-293 cells for library-derived and primary designer leads. Chimeric and humanized 24C05, isotype control IgG1 and 15G11-DI1 to DI11 clones in human IgG1 format were examined for specific binding on human (FIG. 10A) and rhesus (FIG. 10B) ERBB3-transfected HEK-293 cells. IgGs were tested at concentrations ranging from 0.008-500 nM. Concentration-dependent binding was observed against both human and rhesus transfected cells for all ERBB3-specific antibodies but not isotype controls.
[0091] FIG. 11. Development risk ELISAs. Chimeric and humanized 24C05, isotype control IgG1, clinical-stage control antibodies, and 15G11-DI1 to DI11 clones in human IgG1 format were examined for nonspecific binding to the negatively charged biomolecules Insulin and double-stranded DNA (dsDNA). All lead clones demonstrated binding scores below 10 (15G11-DI10 being an exception), significantly lower than either of the negative control IgG1 Ustekinumab and Bevacizumab analogs. Strong off-target binding to insulin or dsDNA, as observed for Bococizumab and Briakinumab analogues, has been shown to be a high-risk indicator of poor pharmacokinetics of therapeutic antibodies.
[0092] FIG. 12A-FIG. 12G. Cell-based ErbB2-ErbB3 antagonism assay. Chimeric and humanized 24C05, isotype control IgG1 and clones 15G11 (FIG. 12A), 16B09 (FIG. 12B), 15G11-DI5 (FIG. 12C), 15G11-DI6 (FIG. 12D), 15G11-DI7 (FIG. 12E), 15G11-DI8 (FIG. 12F), and 15G11-DI9 (FIG. 12G) in human IgG1 format were titrated in a human ErbB3 signaling reporter assay (DiscoverX PathHunter eXpress ErbB2-ErbB3 assay, performed according to manufacturer's instructions). All clones other than the Isotype control induced strong, concentration-dependent ErbB3 antagonism, with highly similar potencies to h24C05.
[0093] FIG. 13. Cell-based ErbB2-ErbB3 antagonism assay for 15G11-DI9. Chimeric and humanized 24C05, isotype control IgG1 and clone 15G11-D19 in human IgG1 format were titrated in a human ErbB3 signaling reporter assay (DiscoverX PathHunter eXpress ErbB2-ErbB3 assay, performed according to manufacturer's instructions). All clones other than the Isotype control induced strong, concentration-dependent ErbB3 antagonism, as evidenced by fold inhibition of signal.
[0094] FIG. 14A-FIG. 14E. T cell epitope peptide content in lead antibody v-domains. The v-domains of 15G11-DI5 (FIG. 14A), 15G11-DI6 (FIG. 14B), 15G11-DI7 (FIG. 14C), 15G11-DI8 (FIG. 14D) and 15G11-DI9 (FIG. 14E) antibodies were examined for the presence of Germline (GE), High Affinity Foreign (HAF), Low Affinity Foreign (LAF) and TCED+ T cell receptor epitopes. In all lead clones, the high-risk epitope content was progressively reduced, and germline epitope content maintained from 15G11-DI5 to 15G11-DI9, with 15G11-DI9 containing no predicted foreign epitopes at all, coupled with high GE content, suggesting that this clone may be fully non-immunogenic in man.
DETAILED DESCRIPTION OF THE INVENTION
[0095] According to a first aspect of the invention, there is provided an antibody molecule which specifically binds to human ERBB3 and optionally also to rhesus monkey ERBB3, or an antigen-binding portion thereof, wherein the antibody molecule or antigen-binding portion comprises a heavy chain variable region with:
[0096] an HCDR1 having amino acids in sequence in the following order: G-F-T-F-S-D-Y-G or any amino acid (such as S)-M-S (SEQ ID NO:1);
[0097] an HCDR2 having amino acids in sequence in the following order: V-S-T-I-S-D-G or any amino acid (such as S, D)-G-T or a conservative substitution of T (such as S)-Y or any amino acid (such as T)-T or any amino acid (such as I)-Y-Y-P or any amino acid (such as A)-D-N or a conservative substitution of N (such as S)-V-K-G (SEQ ID NO:2); and
[0098] an HCDR3 having amino acids in sequence in the following order: E or any amino acid (such as M)-W or any amino acid (such as F, L, M, Q or Y)-G-D-Y or any amino acid (such as A, D, E, H, L, M, N, Q, S, T or VV)-D-G-F or any amino acid (such as I, L, W, Y)-D-Y or any amino acid (such as A, D, E, F, H, I, K, L, M, N, Q, R, S, V, W) (SEQ ID NO:3).
[0099] In some aspects an anti-ERBB3 antibody or antigen-binding portion provided herein specifically binds to a ERBB3 protein comprising or consisting of SEQ ID NO:246 or SEQ ID NO:247. In some aspects an anti-ERBB3 antibody or antigen-binding portion provided herein specifically binds to a ERBB3 protein having an amino acid sequence that is at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to SEQ ID NO:246 or SEQ ID NO:247.
[0100] In aspects of the invention, the HCDR1 of the antibody molecule or antigen-binding portion may exclude the sequence GFTFSDYAMS (SEQ ID NO:4; 24C05 murine/humanized antibody HCDR1 disclosed in WO2011136911A2; US20110256154A1), the HCDR2 of the antibody molecule or antigen-binding portion may exclude the sequence VSTISDGGTYTYYPDNVKG (SEQ ID NO:5; 24C05 murine/humanized antibody HCDR2 disclosed in WO2011136911A2; US20110256154A1), and/or the HCDR3 of the antibody molecule or antigen-binding portion may exclude the sequence EWGDYDGFDY (SEQ ID NO:6; 24C05 murine/humanized antibody HCDR3 disclosed in WO2011136911A2; US20110256154A1).
[0101] The antibody molecule or antigen-binding portion may further comprise a light chain variable region with:
[0102] an LCDR1 having amino acids in sequence in the following order: R-A-S-Q-E or any amino acid (such as S, I, N)-I-S-G or a conservative substitution of G (such as S, T)-Y-L-S or a conservative substitution of S (such as N) (SEQ ID NO:7);
[0103] an LCDR2 having amino acids in sequence in the following order: A or any amino acid (such as E)-A-S-T or a conservative substitution of T (such as S, N)-L-D or any amino acid (such as H, K, Q)-S or T (SEQ ID NO:8); and
[0104] an LCDR3 having amino acids in sequence in the following order: L or any amino acid (such as Q)-Q-Y or any amino acid (such as S)-D or any amino acid (such as Y)-S-Y or any amino acid (such as T, S)-P or any amino acid (such as H)-Y or any amino acid (such as L)-T (SEQ ID NO:9).
[0105] In aspects of the invention, the LCDR1 of the antibody molecule or antigen-binding portion may exclude the sequence RASQEISGYLS (SEQ ID NO:10; 24C05 murine/humanized antibody LCDR1 disclosed in WO2011136911A2; US20110256154A1), and/or the LCDR2 of the antibody molecule or antigen-binding portion may exclude the sequence AASTLDS (SEQ ID NO:11; 24C05 murine/humanized antibody LCDR2 disclosed in WO2011136911A2; US20110256154A1), and/or the LCDR3 of the antibody molecule or antigen-binding portion may exclude the sequence LQYDSYPYT (SEQ ID NO:12; 24C05 murine/humanized antibody LCDR3 disclosed in WO2011136911A2; US20110256154A1).
[0106] In some aspects, disclosed herein is an anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody comprises a heavy chain variable (VH) region and a light chain variable (VL) region, wherein
[0107] (a) the HCDR1 comprises the amino acid sequence G-F-T-F-S-D-Y-X.sub.1-M-S, wherein X.sub.1 is G or any other amino acid (for example, S) (SEQ ID NO:1);
[0108] (b) the HCDR2 comprises V-S-T-I-S-D-X.sub.1-G-X.sub.2-X.sub.3-X.sub.4-Y-Y-X.sub.5-D-X.sub.6-V-K-G- , wherein X.sub.1 is G or any other amino acid (for example, S or D), X.sub.2 is T or a conservative substitution of T (for example, S), X.sub.3 is Y or any other amino acid (for example, T), X.sub.4 is T or any other amino acid (for example, I), X.sub.5 is P or any other amino acid (for example, A), and X.sub.6 is N or a conservative substitution of N (for example, S) (SEQ ID NO:2);
[0109] (c) the HCDR3 comprises X.sub.1-X.sub.2-G-D-X.sub.3-D-G-X.sub.4-D-X.sub.5, wherein X.sub.1 is E or any other amino acid (for example, M), X.sub.2 is W or any other amino acid (for example, F, L, M, Q or Y), X.sub.3 is Y or any other amino acid (for example, A, D, E, H, L, M, N, Q, S, T or W), X.sub.4 is F or any other amino acid (for example, I, L, W or Y), and X.sub.5 is Y or any other amino acid (for example, A, D, E, F, H, I, K, L, M, N, Q, R, S, V or W) (SEQ ID NO:3);
[0110] (d) the LCDR1 comprises R-A-S-Q-X.sub.1-I-S-X.sub.2-Y-L-X.sub.3, wherein X.sub.1 is E or any other amino acid (for example, S, I or N), X.sub.2 is G or a conservative substitution of G (for example, S or T), and X.sub.3 is S or a conservative substitution of S (for example, N) (SEQ ID NO:7);
[0111] (e) the LCDR2 comprises X.sub.1-A-S-X.sub.2-L-X.sub.3-S, wherein X.sub.1 is A or any other amino acid (for example, E), X.sub.2 is T or a conservative substitution of T (for example, S or N), and X.sub.3 is or D any other amino acid (for example, H, K or Q) (SEQ ID NO:8); and
[0112] (f) the LCDR3 comprises X.sub.1-Q-X.sub.2-X.sub.3-S-X.sub.4-X.sub.5-X.sub.6-T, wherein X.sub.1 is L or any other amino acid (for example, Q), X.sub.2 is Y or any other amino acid (for example, S), X.sub.3 is D or any other amino acid (for example, Y), X.sub.4 is Y or any other amino acid (for example, T or S), X.sub.5 is P or any other amino acid (for example, H), and X.sub.6 is Y or any other amino acid (for example, L) (SEQ ID NO:9). In some aspects, the LCDR2 comprises X.sub.1-A-S-X.sub.2-L-X.sub.3-S (SEQ ID NO:8), wherein the seventh residue in the sequence is a conservative substitution of S (for example, T).
[0113] In some aspects, disclosed herein is an anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody comprises a heavy chain variable (VH) region comprising, in amino-terminal to carboxyl-terminal order, FR1-HCDR1-FR2-HCDR2-FR3-HCDR3-FR4 and a light chain variable (VL) region comprising, in amino-terminal to carboxyl-terminal order, FR1-LCDR1-FR2-LCDR2-FR3-LCDR3-FR4, wherein the HCDR1 is SEQ ID NO:1, the HCDR2 is SEQ ID NO:2, the HCDR3 is SEQ ID NO:3, the LCDR1 is SEQ ID NO:7, the LCDR2 is SEQ ID NO:8 and the LCDR3 is SEQ ID NO:9, wherein the heavy chain FR1, FR2, FR3 and FR4 amino acid sequences are the heavy chain FR1, FR2, FR3 and FR4 amino acid sequences in SEQ ID NO: 86 (see Table 2) and wherein the light chain FR1, FR2, FR3 and FR4 amino acid sequences are the light chain FR1, FR2, FR3 and FR4 amino acid sequences in SEQ ID NO: 88 (see Table 2).
[0114] As elaborated herein, the present inventors have succeeded for the first time in generating a number of optimized anti-ERBB3 antibody molecules using CDR sequences derived from the murine anti-ERBB3 antibody 24C05 disclosed in WO2011136911A2; US20110256154A1. In embodiments of the present invention, these antibody molecules have been selected to have binding specificity to both human ERBB3 as well as rhesus monkey ERBB3 (to facilitate in vivo studies in an appropriate animal test species). Further refining of the optimized antibody molecules as described herein has provided improved variable domain stability, higher expression yields, and/or reduced immunogenicity.
[0115] Preferred optimized anti-ERBB3 antibody molecules of the present invention do not necessarily have the maximum number of human germline substitutions at corresponding murine CDR or other (such as framework) amino acid positions. As elaborated in the experimental section below, we have found that "maximally humanized" antibody molecules are not necessary "maximally optimized" in terms of anti-ERBB3 binding characteristics and/or other desirable features.
[0116] The present invention encompasses modifications to the amino acid sequence of the antibody molecule or antigen-binding portion thereof as defined herein. For example, the invention includes antibody molecules and corresponding antigen-binding portions thereof comprising functionally equivalent variable regions and CDRs which do not significantly affect their properties as well as variants which have enhanced or decreased activity and/or affinity. For example, the amino acid sequence may be mutated to obtain an antibody with the desired binding affinity to ERBB3. Insertions which include amino- and/or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing a hundred or more residues, as well as intrasequence insertions of single or multiple amino acid residues, are envisaged. Examples of terminal insertions include an antibody molecule with an N-terminal methionyl residue or the antibody molecule fused to an epitope tag. Other insertional variants of the antibody molecule include the fusion to the N- or C-terminus of the antibody of an enzyme or a polypeptide which increases the half-life of the antibody in the blood circulation.
[0117] The antibody molecule or antigen-binding portion of the invention may include glycosylated and nonglycosylated polypeptides, as well as polypeptides with other post-translational modifications, such as, for example, glycosylation with different sugars, acetylation, and phosphorylation. The antibody molecule or antigen-binding portion of the invention may be mutated to alter such post-translational modifications, for example by adding, removing or replacing one or more amino acid residues to form or remove a glycosylation site.
[0118] The antibody molecule or antigen-binding portion of the invention may be modified for example by amino acid substitution to remove potential proteolytic sites in the antibody.
[0119] In the antibody molecule or antigen-binding portion thereof, the HCDR1 may have the amino acid sequence: G-F-T-F-S-D-Y-E/G/H/N/R/S/T/Q/V-M-S (SEQ ID NO:38); the HCDR2 may have the amino acid sequence: V-S-T-I-S-D-G/S/D-G-T/S-Y/T-T/I-Y-Y-P/A-D-N/S-V-K-G (SEQ ID NO:39); and the HCDR3 may have the amino acid sequence: E/M-W/F/L/M/Q/Y-G-D-Y/A/D/E/H/UM/N/Q/S/T/W-D-G-F/I/L/W/Y-D-Y/A/D/E/F/H/I/- K/UM/N/Q/R/S/V/W (SEQ ID NO:40).
[0120] For example, the HCDR1 may have the amino acid sequence: G-F-T-F-S-D-Y-G/S-M-S (SEQ ID NO:41); the HCDR2 may have the amino acid sequence: V-S-T-1-S-D-G/S-G-S-Y/T-T/I-Y-Y-P/A-D-S-V-K-G (SEQ ID NO:42); and the HCDR3 may have the amino acid sequence: E-W/L/Y-G-D-Y-D-G-F-D-Y/E/F/H/N (SEQ ID NO:43).
[0121] In the antibody molecule or antigen-binding portion thereof, the LCDR1 may have the amino acid sequence: R-A-S-Q-E/S/I/N-I-S-G/S-Y-L-S/N (SEQ ID NO:44); the LCDR2 may have the amino acid sequence: A/E-A-S-T/S/N-L-D/H/K/Q-S (SEQ ID NO:45); and the LCDR3 may have the amino acid sequence: L/Q-Q-Y/S-D/Y-S-Y/T-P/H-Y/L-T (SEQ ID NO:46). In the antibody molecule or antigen-binding portion thereof, the LCDR1 may have the amino acid sequence: R-A-S-Q-E/S/I/N-I-S-G/S/T-Y-L-S/N; the LCDR2 may have the amino acid sequence: A/E-A-S-T/S/N-L-D/H/K/Q-S/T; and the LCDR3 may have the amino acid sequence: L/Q-Q-Y/S-D/Y-S-Y/T/S-P/H-Y/L-T.
[0122] For example, the LCDR1 may have the amino acid sequence: R-A-S-Q-E/S-I-S-G/S-Y-L-S/N (SEQ ID NO:47); the LCDR2 may have the amino acid sequence: A-A-S-T/S-L-D/Q-S (SEQ ID NO:48); and the LCDR3 may have the amino acid sequence: L-Q-Y-D/Y-S-T-P-Y/L-T (SEQ ID NO:49). For example, the LCDR1 may have the amino acid sequence: R-A-S-Q-E/S-I-S-G/S/T-Y-L-S/N; the LCDR2 may have the amino acid sequence: A-A-S-T/S-L-D/Q-S/T; and the LCDR3 may have the amino acid sequence: L-Q-Y-D/Y-S-T/S-P-Y/L-T.
[0123] In specific embodiments of the invention, the antibody molecule or antigen-binding portion may comprise:
[0124] (a) the amino acid sequences RASQSISSYLS (SEQ ID NO:16; LCDR1), AASTLQS (SEQ ID NO:26; LCDR2), LQYDSTPLT (SEQ ID NO:18; LCDR3), GFTFSDYGMS (SEQ ID NO:13; HCDR1), VSTISDGGSYTYYADNVKG (SEQ ID NO:31; HCDR2), EWGDYDGFDF (SEQ ID NO:15; HCDR3), [Clone 15D10]; or
[0125] (b) the amino acid sequences RASQSISGYLS (SEQ ID NO:30; LCDR1), AASTLQS (SEQ ID NO:26; LCDR2), LQYDSTPYT (SEQ ID NO:23; LCDR3), GFTFSDYGMS (SEQ ID NO:13; HCDR1), VSTISDGGSYTYYADSVKG (SEQ ID NO:28; HCDR2), EWGDYDGFDE (SEQ ID NO:29; HCDR3), [Clone 17H10]; or
[0126] (c) the amino acid sequences RASQSISSYLN (SEQ ID NO:50; LCDR1), AASSLDS (SEQ ID NO:22; LCDR2), LQYDSTPLT (SEQ ID NO:18; LCDR3), GFTFSDYGMS (SEQ ID NO:13; HCDR1), VSTISDGGSYTYYADSVKG (SEQ ID NO:28; HCDR2), EYGDYDGFDY (SEQ ID NO:51; HCDR3), [Clone 09D12]; or
[0127] (d) the amino acid sequences RASQEISSYLS (SEQ ID NO:21; LCDR1), AASSLQS (SEQ ID NO:17; LCDR2), LQYDSTPLT (SEQ ID NO:18; LCDR3), GFTFSDYGMS (SEQ ID NO:13; HCDR1), VSTISDSGSYIYYADSVKG (SEQ ID NO:14; HCDR2), EWGDYDGFDH (SEQ ID NO:27; HCDR3), [Clone 15D03]; or
[0128] (e) the amino acid sequences RASQIISSYLS (SEQ ID NO:52; LCDR1), AASSLDS (SEQ ID NO:22; LCDR2), LQYYSTPLT (SEQ ID NO:53; LCDR3), GFTFSDYGMS (SEQ ID NO:13; HCDR1), VSTISDSGSYTYYADSVKG (SEQ ID NO:54; HCDR2), EWGDYDGFDN (SEQ ID NO:55; HCDR3), [Clone 11H02]; or
[0129] (f) the amino acid sequences RASQEISSYLS (SEQ ID NO:21; LCDR1), AASSLDS (SEQ ID NO:22; LCDR2), LQYDSTPYT (SEQ ID NO:23; LCDR3), GFTFSDYGMS (SEQ ID NO:13; HCDR1), VSTISDSGSYTYYPDSVKG (SEQ ID NO:19; HCDR2), ELGDYDGFDY (SEQ ID NO:20; HCDR3), [Clone 15G11]; or
[0130] (g) the amino acid sequences RASQSISSYLS (SEQ ID NO:16; LCDR1), AASSLQS (SEQ ID NO:17; LCDR2), LQYDSTPLT (SEQ ID NO:18; LCDR3), GFTFSDYGMS (SEQ ID NO:13; HCDR1), VSTISDSGTTIYYADNVKG (SEQ ID NO:56; HCDR2), EYGDYDGFDY (SEQ ID NO:51; HCDR3), [Clone 15E02]; or
[0131] (h) the amino acid sequences RASQSISSYLS (SEQ ID NO:16; LCDR1), AASSLQS (SEQ ID NO:17; LCDR2), LQYDSTPLT (SEQ ID NO:18; LCDR3), GFTFSDYSMS (SEQ ID NO:24; HCDR1), VSTISDGGSYTYYPDSVKG (SEQ ID NO:57; HCDR2), ELGDYDGFDY (SEQ ID NO:20; HCDR3), [Clone 09H02]; or
[0132] (i) the amino acid sequences RASQEISSYLS (SEQ ID NO:21; LCDR1), AASTLQS (SEQ ID NO:26; LCDR2), LQYDSTPLT (SEQ ID NO:18; LCDR3), GFTFSDYSMS (SEQ ID NO:24; HCDR1), VSTISDSGTYTYYPDSVKG (SEQ ID NO:25; HCDR2), EWGDYDGFDF (SEQ ID NO:15; HCDR3), [Clone 16609]; or
[0133] (j) the amino acid sequences RASQSISSYLS (SEQ ID NO:16; LCDR1), AASSLQS (SEQ ID NO:17; LCDR2), LQYDSTPLT (SEQ ID NO:18; LCDR3), GFTFSDYGMS (SEQ ID NO:13; HCDR1), VSTISDSGSYIYYADSVKG (SEQ ID NO:14; HCDR2), ELGDYDGFDY (SEQ ID NO:20; HCDR3), [Clone MH1]; or
[0134] (k) the amino acid sequences RASQSISSYLS (SEQ ID NO:16; LCDR1), AASSLQS (SEQ ID NO:17; LCDR2), LQYDSTPLT (SEQ ID NO:18; LCDR3), GFTFSDYGMS (SEQ ID NO:13; HCDR1), VSTISDSGSYIYYADSVKG (SEQ ID NO:14; HCDR2), EWGDYDGFDF (SEQ ID NO:15; HCDR3), [Clone MH2]; or
[0135] (l) the amino acid sequences RASQSISSYLS (SEQ ID NO:16; LCDR1), AASSLQS (SEQ ID NO:17; LCDR2), LQYDSTPLT (SEQ ID NO:18; LCDR3), GFTFSDYSMS (SEQ ID NO:24; HCDR1), VSTISDSGSTIYYADSVKG (SEQ ID NO:58; HCDR2), EWGDYDGFDF (SEQ ID NO:15; HCDR3), [Clone MH3]; or
[0136] (m) the amino acid sequences RASQSISSYLS (SEQ ID NO:16; LCDR1), AASSLQS (SEQ ID NO:17; LCDR2), LQYDSTPLT (SEQ ID NO:18; LCDR3), GFTFSDYGMS (SEQ ID NO:13; HCDR1), VSTISDSGSYIYYADSVKG (SEQ ID NO:14; HCDR2), EWGDYDGFDE (SEQ ID NO:29; HCDR3), [Clone MH4]; or
[0137] (n) the amino acid sequences RASQSISSYLS (SEQ ID NO:16; LCDR1), AASSLQS (SEQ ID NO:17; LCDR2), LQYDSTPLT (SEQ ID NO:18; LCDR3), GFTFSDYGMS (SEQ ID NO:13; HCDR1), VSTISDSGSTIYYADSVKG (SEQ ID NO:58; HCDR2), EYGDYDGFDY (SEQ ID NO:51; HCDR3), [Clone MH5]; or
[0138] (o) the amino acid sequences RASQSISSYLN (SEQ ID NO:50; LCDR1), AASSLQS (SEQ ID NO:17; LCDR2), LQYDSTPLT (SEQ ID NO:18; LCDR3), GFTFSDYGMS (SEQ ID NO:13; HCDR1), VSTISDSGSTIYYADSVKG (SEQ ID NO:58; HCDR2), EYGDYDGFDY (SEQ ID NO:51; HCDR3), [Clone TTP]; or
[0139] (p) the amino acid sequences RASQEISTYLS (SEQ ID NO: 261; LCDR1), AASTLQS (SEQ ID NO:26; LCDR2), LQYDSSPLT (SEQ ID NO: 262; LCDR3), GFTFSDYSMS (SEQ ID NO: 24; HCDR1), VSTISDSGTYTYYPDSVKG (SEQ ID NO: 25; HCDR2), EWGDYDGFDF (SEQ ID NO: 15; HCDR3), [Clone 15G11-D19]; or
[0140] (q) the amino acid sequences RASQEISSYLS (SEQ ID NO: 21; LCDR1), AASSLDT (SEQ ID NO: 263; LCDR2), LQYDSTPYT (SEQ ID NO: 23; LCDR3), GFTFSDYSMS (SEQ ID NO: 24; HCDR1), VSTISDSGTYTYYPDSVKG (SEQ ID NO: 25; HCDR2), EWGDYDGFDF (SEQ ID NO: 15; HCDR3), [Clone 15G11-D15].
[0141] In some aspects, disclosed herein is an anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody comprises a heavy chain variable (VH) region and a light chain variable (VL) region, wherein
[0142] (a) the VH region amino acid sequence comprises HCDR1 of GFTFSDYSMS (SEQ ID NO: 24), HCDR2 of VSTISDSGTYTYYPDSVKG (SEQ ID NO: 25) and HCDR3 of EWGDYDGFDF (SEQ ID NO: 15); and the VL region amino acid sequence comprises LCDR1 of RASQEISTYLS (SEQ ID NO: 261), LCDR2 of AASTLQS (SEQ ID NO:26) and LCDR3 of LQYDSSPLT (SEQ ID NO: 262);
[0143] (b) the VH region amino acid sequence comprises HCDR1 of GFTFSDYSMS (SEQ ID NO: 24), HCDR2 of VSTISDSGTYTYYPDSVKG (SEQ ID NO: 25) and HCDR3 of EWGDYDGFDF (SEQ ID NO: 15); and the VL region amino acid sequence comprises LCDR1 of RASQEISSYLS (SEQ ID NO: 21), LCDR2 of AASSLDT (SEQ ID NO: 263) and LCDR3 of LQYDSTPYT (SEQ ID NO: 23);
[0144] (c) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDSGSYTYYPDSVKG (SEQ ID NO:19) and HCDR3 of ELGDYDGFDY (SEQ ID NO:20); and the VL region amino acid sequence comprises LCDR1 of RASQEISSYLS (SEQ ID NO:21), LCDR2 of AASSLDS (SEQ ID NO:22) and LCDR3 of LQYDSTPYT (SEQ ID NO:23);
[0145] (d) the VH region amino acid sequence comprises HCDR1 of GFTFSDYSMS (SEQ ID NO:24), HCDR2 of VSTISDSGTYTYYPDSVKG (SEQ ID NO:25) and HCDR3 of EWGDYDGFDF (SEQ ID NO:15); and the VL region amino acid sequence comprises LCDR1 of RASQEISSYLS (SEQ ID NO:21), LCDR2 of AASTLQS (SEQ ID NO:26) and LCDR3 of LQYDSTPLT (SEQ ID NO:18);
[0146] (e) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDSGSYIYYADSVKG (SEQ ID NO:14) and HCDR3 of EWGDYDGFDF (SEQ ID NO:15); and the VL region amino acid sequence comprises LCDR1 of RASQSISSYLS (SEQ ID NO:16), LCDR2 of AASSLQS (SEQ ID NO:17) and LCDR3 of LQYDSTPLT (SEQ ID NO:18);
[0147] (f) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDSGSYIYYADSVKG (SEQ ID NO:14) and HCDR3 of EWGDYDGFDH (SEQ ID NO:27); and the VL region amino acid sequence comprises LCDR1 of RASQEISSYLS (SEQ ID NO:21), LCDR2 of AASSLQS (SEQ ID NO:17) and LCDR3 of LQYDSTPLT (SEQ ID NO:18);
[0148] (g) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDGGSYTYYADSVKG (SEQ ID NO:28) and HCDR3 of EWGDYDGFDE (SEQ ID NO:29); and the VL region amino acid sequence comprises LCDR1 of RASQSISGYLS (SEQ ID NO:30), LCDR2 of AASTLQS (SEQ ID NO:26) and LCDR3 of LQYDSTPYT (SEQ ID NO:23);
[0149] (h) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDGGSYTYYADNVKG (SEQ ID NO:31) and HCDR3 of EWGDYDGFDF (SEQ ID NO:15); and the VL region amino acid sequence comprises LCDR1 of RASQSISSYLS (SEQ ID NO:16), LCDR2 of AASTLQS (SEQ ID NO:26) and LCDR3 of LQYDSTPLT (SEQ ID NO:18); or
[0150] (i) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDSGSYIYYADSVKG (SEQ ID NO:14) and HCDR3 of EWGDYDGFDE (SEQ ID NO:29); and the VL region amino acid sequence comprises LCDR1 of RASQSISSYLS (SEQ ID NO:16), LCDR2 of AASSLQS (SEQ ID NO:17) and LCDR3 of LQYDSTPLT (SEQ ID NO:18).
[0151] In some aspects, disclosed herein is an anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody comprises a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH region comprises any one of the VH region amino acid sequences in Table 7 or 8 and the VL region comprises any one of the VL region amino acid sequences in Table 6 or 8.
[0152] In some aspects, disclosed herein is an anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody comprises a heavy chain variable (VH) region and a light chain variable (VL) region, wherein
[0153] (a) the VH region amino acid sequence comprises SEQ ID NO:236 and the VL region amino acid sequence comprises SEQ ID NO:225;
[0154] (b) the VH region amino acid sequence comprises SEQ ID NO:232 and the VL region amino acid sequence comprises SEQ ID NO:221;
[0155] (c) the VH region amino acid sequence comprises SEQ ID NO:253 and the VL region amino acid sequence comprises SEQ ID NO:254;
[0156] (d) the VH region amino acid sequence comprises SEQ ID NO:255 and the VL region amino acid sequence comprises SEQ ID NO:256;
[0157] (e) the VH region amino acid sequence comprises SEQ ID NO:228 and the VL region amino acid sequence comprises SEQ ID NO:217;
[0158] (f) the VH region amino acid sequence comprises SEQ ID NO:229 and the VL region amino acid sequence comprises SEQ ID NO:218;
[0159] (g) the VH region amino acid sequence comprises SEQ ID NO:230 and the VL region amino acid sequence comprises SEQ ID NO:219;
[0160] (h) the VH region amino acid sequence comprises SEQ ID NO:231 and the VL region amino acid sequence comprises SEQ ID NO:220;
[0161] (i) the VH region amino acid sequence comprises SEQ ID NO:233 and the VL region amino acid sequence comprises SEQ ID NO:222;
[0162] (j) the VH region amino acid sequence comprises SEQ ID NO:234 and the VL region amino acid sequence comprises SEQ ID NO:223;
[0163] (k) the VH region amino acid sequence comprises SEQ ID NO:235 and the VL region amino acid sequence comprises SEQ ID NO:224;
[0164] (l) the VH region amino acid sequence comprises SEQ ID NO:237 and the VL region amino acid sequence comprises SEQ ID NO:226; or
[0165] (m) the VH region amino acid sequence comprises SEQ ID NO:238 and the VL region amino acid sequence comprises SEQ ID NO:227.
[0166] In some aspects, disclosed herein is an anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody comprises a heavy chain variable (VH) region and a light chain variable (VL) region, wherein
[0167] (a) the VH region amino acid sequence is at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to SEQ ID NO:236 and the VL region amino acid sequence is at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to SEQ ID NO:225;
[0168] (b) the VH region amino acid sequence is at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to SEQ ID NO:232 and the VL region amino acid sequence is at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to SEQ ID NO:221;
[0169] (c) the VH region amino acid sequence is at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to SEQ ID NO:253 and the VL region amino acid sequence is at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to SEQ ID NO:254; or
[0170] (d) the VH region amino acid sequence is at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to SEQ ID NO:255 and the VL region amino acid sequence is at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to SEQ ID NO:256.
[0171] In some aspects, the antibody or antigen-binding portion as defined herein may be isolated.
[0172] The antibody molecule or antigen-binding portion as defined herein may cross-compete for binding to ERBB3 with an antibody or antigen-binding portion thereof comprising the sets of CDRs disclosed herein. In some embodiments, the invention provides an isolated anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody or antigen-binding portion cross-competes for binding to ERBB3 with the antibody or antigen-binding portion comprising the sets of CDRs disclosed herein; and (a) comprises fully germline human framework amino acid sequences; and/or (b) does not comprise an isomerization site in the LCDR2; and/or (c) does not comprise a `DG` isomerization site in the HCDR2; and/or (d) does not comprise an oxidation site at position 2 in the HCDR3; and/or and/or (e) exhibits a reduced number of predicted foreign human T cell receptor binding peptides in its v-domains in comparison to h24C05; and/or (f) contains no predicted foreign human T cell receptor binding peptides in its v-domains. In some embodiments, the anti-ERBB3 antibody or an antigen-binding portion thereof does not comprise an oxidation site at position 2 (e.g., W) in the HCDR3.
[0173] The terms "cross-compete", "cross-competition", "cross-block", "cross-blocked" and "cross-blocking" are used interchangeably herein to mean the ability of an antibody or portion thereof to interfere with the binding directly or indirectly through allosteric modulation of the anti-ERBB3 antibodies of the invention to the target ERBB3 (e.g., human ERBB3). The extent to which an antibody or portion thereof is able to interfere with the binding of another to the target, and therefore whether it can be said to cross-block or cross-compete according to the invention, can be determined using competition binding assays. One example of a binding competition assay is Homogeneous Time Resolved Fluorescence (HTRF). One particularly suitable quantitative cross-competition assay uses a FACS- or an AlphaScreen-based approach to measure competition between the labelled (e.g. His tagged, biotinylated or radioactive labelled) antibody or portion thereof and the other antibody or portion thereof in terms of their binding to the target. In general, a cross-competing antibody or portion thereof is, for example, one which will bind to the target in the cross-competition assay such that, during the assay and in the presence of a second antibody or portion thereof, the recorded displacement of the immunoglobulin single variable domain or polypeptide according to the invention is up to 100% (e.g. in a FACS based competition assay) of the maximum theoretical displacement (e.g. displacement by cold (e.g. unlabeled) antibody or fragment thereof that needs to be cross-blocked) by the potentially cross-blocking antibody or fragment thereof that is present in a given amount. Preferably, cross-competing antibodies or portions thereof have a recorded displacement that is between 10% and 100%, or between 50% and 100%.
[0174] The antibody molecule or antigen-binding portion as defined herein may comprise one or more substitutions, deletions and/or insertions which remove a post-translational modification (PTM) site, for example a glycosylation site (N-linked or O-linked), a deamination site, a phosphorylation site or an isomerisation/fragmentation site.
[0175] More than 350 types of PTM are known. Key forms of PTM include phosphorylation, glycosylation (N- and O-linked), sumoylation, palmitoylation, acetylation, sulfation, myristoylation, prenylation and methylation (of K and R residues). Statistical methods to identify putative amino acid sites responsible for specific PTMs are well known in the art (see Zhou et al., 2016, Nature Protocols 1: 6588-1321). Removal of such a site for example by substitution, deletion and/or insertion and then optionally testing (experimentally and/or theoretically) for (a) binding activity and/or (b) loss of the PTM is contemplated.
[0176] For example, the 24C05 murine LCDR2 (as defined herein, i.e. the amino acid sequence AASTLDS (SEQ ID NO:11)) has been identified to have a putative isomerisation site at residue 6. Removal this site at equivalent positions in an LCDR2 of the invention, for example by substitution of D (such as to Q), is envisaged (as for example in clone MH2 and others found in Tables 3 and 4).
[0177] In a further example, the 24C05 murine HCDR2 (as defined herein, i.e. the amino acid sequence VSTISDGGTYTYYPDNVKG (SEQ ID NO:5)) has been identified to have a putative isomerisation site at residue 6 (D). Reduction in chemical modification risk this site at equivalent positions in an HCDR2 of the invention, for example by substitution of G (such as to S, or D), is envisaged (as for example in clone 15G11 and others found in Tables 3 and 4).
[0178] In a further example, the 24C05 murine HCDR3 (as defined herein, i.e. the amino acid sequence EWGDYDGFDY (SEQ ID NO:6)) has been identified to have a putative oxidation site at residue 2 (VV). Removal this site at equivalent positions in an LCDR1 of the invention, for example by substitution of W (such as to L or Y), is envisaged (as for example in clone 15G11 and others found in Tables 3 and 4).
[0179] The antibody molecule or antigen-binding portion thereof may be human, humanized or chimeric.
[0180] The antibody molecule or antigen-binding portion thereof may comprise one or more human variable domain framework scaffolds into which the CDRs have been inserted. For example, the VH region, the VL region, or both the VH and the VL region may comprise one or more human framework region amino acid sequences.
[0181] The antibody molecule or antigen-binding portion thereof may comprise an IGHV3-11 human germline scaffold into which the corresponding HCDR sequences have been inserted. The antibody molecule or antigen-binding portion thereof may comprise a VH region that comprises an IGHV3-11 human germline scaffold amino acid sequence into which a set of corresponding HCDR1, HCDR2 and HCDR3 amino acid sequences have been inserted.
[0182] The antibody molecule or antigen-binding portion thereof may comprise an IGKV1-39 human germline scaffold into which the corresponding LCDR sequences have been inserted. The antibody molecule or antigen-binding portion thereof may comprise a VL region that comprises an IGKV1-39 human germline scaffold amino acid sequence into which a set of corresponding LCDR1, LCDR2 and LCDR3 amino acid sequences have been inserted.
[0183] The antibody molecule or antigen-binding portion thereof may comprise an IGHV3-11 human germline scaffold into which the corresponding HCDR sequences have been inserted and an IGKV1-39 human germline scaffold into which the corresponding LCDR sequences have been inserted. The antibody molecule or antigen-binding portion thereof may comprise a VH region that comprises an IGHV3-11 human germline scaffold amino acid sequence into which a set of corresponding HCDR1, HCDR2 and HCDR3 amino acid sequences have been inserted and a VL region that comprises an IGKV1-39 human germline scaffold amino acid sequence into which a set of corresponding LCDR1, LCDR2 and LCDR3 amino acid sequences have been inserted. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 amino acid sequences may be the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 amino acid sequences of any one of the clones in Table 4 (with all six CDR sequences being from the same clone).
[0184] In some aspects, the antibody molecule or antigen-binding portion thereof may comprise an immunoglobulin constant region. In some embodiments, the immunoglobulin constant region is IgG1, IgG2, IgG3, IgG4, IgA1 or IgA2. In additional embodiments, the immunoglobulin constant region is IgG1, IgG2, IgG3, IgG1null, IgG4(S228P), IgA1 or IgA2. The antibody molecule or antigen-binding portion thereof may comprise an immunologically inert constant region. In some aspects, an anti-ERBB3 antibody or antigen-binding portion thereof may comprise an immunoglobulin constant region comprising a wild-type human IgG1 constant region, a human IgG1 constant region comprising the amino acid substitutions L234A, L235A and G237A or a human IgG1 constant region comprising the amino acid substitutions L234A, L235A, G237A and P331S. In some aspects, an anti-ERBB3 antibody or antigen-binding portion thereof may comprise an immunoglobulin constant region comprising a wild-type human IgG2 constant region or a wild-type human IgG4 constant region. In some aspects, an anti-ERBB3 antibody may comprise an immunoglobulin constant region comprising any one of the amino acid sequences in Table 9. The Fc region sequences in Table 9 begin at the CH1 domain. In some aspects, an anti-ERBB3 antibody may comprise an immunoglobulin constant region comprising an amino acid sequence of an Fc region of human IgG4, human IgG4(S228P), human IgG2, human IgG1, human IgG1-3M or human IgG1-4M. For example, the human IgG4(S228P) Fc region comprises the following substitution compared to the wild-type human IgG4 Fc region: S228P. For example, the human IgG1-3M Fc region comprises the following substitutions compared to the wild-type human IgG1 Fc region: L234A, L235A and G237A, while the human IgG1-4M Fc region comprises the following substitutions compared to the wild-type human IgG1 Fc region: L234A, L235A, G237A and P331S. In some aspects, a position of an amino acid residue in a constant region of an immunoglobulin molecule is numbered according to EU nomenclature (Ward et al., 1995 Therap. Immunol. 2:77-94). In some aspects, an immunoglobulin constant region may comprise an RDELT (SEQ ID NO:248) motif or an REEM (SEQ ID NO:249) motif (underlined in Table 9). The REEM (SEQ ID NO:249) allotype is found in a smaller human population than the RDELT (SEQ ID NO:248) allotype. In some aspects, an anti-ERBB3 antibody may comprise an immunoglobulin constant region comprising any one of SEQ ID NOS:239-245. In some aspects, an anti-ERBB3 antibody may comprise the six CDR amino acid sequences of any one of the clones in Table 4 and any one of the Fc region amino acid sequences in Table 9. In some aspects, an anti-ERBB3 antibody may comprise an immunoglobulin heavy chain constant region comprising any one of the Fc region amino acid sequences in Table 9 and an immunoglobulin light chain constant region that is a kappa light chain constant region or a lambda light chain constant region.
[0185] In some aspects, disclosed herein is an anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody comprises a heavy chain variable (VH) region, a light chain variable (VL) region and a heavy chain constant region, wherein
[0186] (a) the VH region amino acid sequence comprises HCDR1 of GFTFSDYSMS (SEQ ID NO: 24), HCDR2 of VSTISDSGTYTYYPDSVKG (SEQ ID NO: 25) and HCDR3 of EWGDYDGFDF (SEQ ID NO: 15); the VL region amino acid sequence comprises LCDR1 of RASQEISTYLS (SEQ ID NO: 261), LCDR2 of AASTLQS (SEQ ID NO:26) and LCDR3 of LQYDSSPLT (SEQ ID NO: 262); and the heavy chain constant region comprises any one of SEQ ID NOS: 239-245;
[0187] (b) the VH region amino acid sequence comprises HCDR1 of GFTFSDYSMS (SEQ ID NO: 24), HCDR2 of VSTISDSGTYTYYPDSVKG (SEQ ID NO: 25) and HCDR3 of EWGDYDGFDF (SEQ ID NO: 15); the VL region amino acid sequence comprises LCDR1 of RASQEISSYLS (SEQ ID NO: 21), LCDR2 of AASSLDT (SEQ ID NO: 263) and LCDR3 of LQYDSTPYT (SEQ ID NO: 23); and the heavy chain constant region comprises any one of SEQ ID NOS: 239-245;
[0188] (c) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDSGSYTYYPDSVKG (SEQ ID NO:19) and HCDR3 of ELGDYDGFDY (SEQ ID NO:20); the VL region amino acid sequence comprises LCDR1 of RASQEISSYLS (SEQ ID NO:21), LCDR2 of AASSLDS (SEQ ID NO:22) and LCDR3 of LQYDSTPYT (SEQ ID NO:23); and the heavy chain constant region comprises any one of SEQ ID NOS: 239-245;
[0189] (d) the VH region amino acid sequence comprises HCDR1 of GFTFSDYSMS (SEQ ID NO:24), HCDR2 of VSTISDSGTYTYYPDSVKG (SEQ ID NO:25) and HCDR3 of EWGDYDGFDF (SEQ ID NO:15); the VL region amino acid sequence comprises LCDR1 of RASQEISSYLS (SEQ ID NO:21), LCDR2 of AASTLQS (SEQ ID NO:26) and LCDR3 of LQYDSTPLT (SEQ ID NO:18); and the heavy chain constant region comprises any one of SEQ ID NOS: 239-245;
[0190] (e) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDSGSYIYYADSVKG (SEQ ID NO:14) and HCDR3 of EWGDYDGFDF (SEQ ID NO:15); the VL region amino acid sequence comprises LCDR1 of RASQSISSYLS (SEQ ID NO:16), LCDR2 of AASSLQS (SEQ ID NO:17) and LCDR3 of LQYDSTPLT (SEQ ID NO:18); and the heavy chain constant region comprises any one of SEQ ID NOS: 239-245;
[0191] (f) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDSGSYIYYADSVKG (SEQ ID NO:14) and HCDR3 of EWGDYDGFDH (SEQ ID NO:27); the VL region amino acid sequence comprises LCDR1 of RASQEISSYLS (SEQ ID NO:21), LCDR2 of AASSLQS (SEQ ID NO:17) and LCDR3 of LQYDSTPLT (SEQ ID NO:18); and the heavy chain constant region comprises any one of SEQ ID NOS: 239-245;
[0192] (g) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDGGSYTYYADSVKG (SEQ ID NO:28) and HCDR3 of EWGDYDGFDE (SEQ ID NO:29); the VL region amino acid sequence comprises LCDR1 of RASQSISGYLS (SEQ ID NO:30), LCDR2 of AASTLQS (SEQ ID NO:26) and LCDR3 of LQYDSTPYT (SEQ ID NO:23); and the heavy chain constant region comprises any one of SEQ ID NOS: 239-245;
[0193] (h) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDGGSYTYYADNVKG (SEQ ID NO:31) and HCDR3 of EWGDYDGFDF (SEQ ID NO:15); the VL region amino acid sequence comprises LCDR1 of RASQSISSYLS (SEQ ID NO:16), LCDR2 of AASTLQS (SEQ ID NO:26) and LCDR3 of LQYDSTPLT (SEQ ID NO:18); and the heavy chain constant region comprises any one of SEQ ID NOS: 239-245; or
[0194] (i) the VH region amino acid sequence comprises HCDR1 of GFTFSDYGMS (SEQ ID NO:13), HCDR2 of VSTISDSGSYIYYADSVKG (SEQ ID NO:14) and HCDR3 of EWGDYDGFDE (SEQ ID NO:29); the VL region amino acid sequence comprises LCDR1 of RASQSISSYLS (SEQ ID NO:16), LCDR2 of AASSLQS (SEQ ID NO:17) and LCDR3 of LQYDSTPLT (SEQ ID NO:18); and the heavy chain constant region comprises any one of SEQ ID NOS: 239-245.
[0195] In some aspects, disclosed herein is an anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody comprises a heavy chain variable (VH) region, a light chain variable (VL) region and a heavy chain constant region, wherein
[0196] (a) the VH region amino acid sequence comprises or consists of SEQ ID NO:236; the VL region amino acid sequence comprises or consists of SEQ ID NO:225; and the heavy chain constant region comprises a wild-type human IgG4 constant region, a human IgG4 constant region comprising the amino acid substitution S228P, a wild-type human IgG2 constant region; a wild-type human IgG1 constant region or a human IgG1 constant region comprising the amino acid substitutions L234A, L235A and G237A;
[0197] (b) the VH region amino acid sequence comprises or consists of SEQ ID NO:232; the VL region amino acid sequence comprises or consists of SEQ ID NO:221; and the heavy chain constant region comprises a wild-type human IgG4 constant region, a human IgG4 constant region comprising the amino acid substitution S228P, a wild-type human IgG2 constant region; a wild-type human IgG1 constant region or a human IgG1 constant region comprising the amino acid substitutions L234A, L235A and G237A;
[0198] (c) the VH region amino acid sequence comprises or consists of SEQ ID NO:253; the VL region amino acid sequence comprises or consists of SEQ ID NO:254; and the heavy chain constant region comprises a wild-type human IgG4 constant region, a human IgG4 constant region comprising the amino acid substitution S228P, a wild-type human IgG2 constant region; a wild-type human IgG1 constant region or a human IgG1 constant region comprising the amino acid substitutions L234A, L235A and G237A; or
[0199] (d) the VH region amino acid sequence comprises or consists of SEQ ID NO:255; the VL region amino acid sequence comprises or consists of SEQ ID NO:256; and the heavy chain constant region comprises a wild-type human IgG4 constant region, a human IgG4 constant region comprising the amino acid substitution S228P, a wild-type human IgG2 constant region; a wild-type human IgG1 constant region or a human IgG1 constant region comprising the amino acid substitutions L234A, L235A and G237A.
[0200] In some aspects, disclosed herein is an anti-ERBB3 antibody or an antigen-binding portion thereof, wherein the antibody comprises a heavy chain variable (VH) region, a light chain variable (VL) region and a heavy chain constant region, wherein
[0201] (a) the VH region amino acid sequence comprises or consists of SEQ ID NO:236; the VL region amino acid sequence comprises or consists of SEQ ID NO:225; and the heavy chain constant region comprises any one of SEQ ID NOS: 239-245;
[0202] (b) the VH region amino acid sequence comprises or consists of SEQ ID NO:232; the VL region amino acid sequence comprises or consists of SEQ ID NO:221; and the heavy chain constant region comprises any one of SEQ ID NOS: 239-245;
[0203] (c) the VH region amino acid sequence comprises or consists of SEQ ID NO:253; the VL region amino acid sequence comprises or consists of SEQ ID NO:254; and the heavy chain constant region comprises any one of SEQ ID NOS: 239-245; or
[0204] (d) the VH region amino acid sequence comprises or consists of SEQ ID NO:255; the VL region amino acid sequence comprises or consists of SEQ ID NO:256; and the heavy chain constant region comprises any one of SEQ ID NOS: 239-245.
[0205] In some aspects, an anti-ERBB3 antibody may be immune effector null. In some aspects, an anti-ERBB3 antibody or an antigen-binding portion thereof does not induce immune effector function and, optionally, suppresses immune effector function. In some aspects, an anti-ERBB3 antibody may lack measurable binding to human Fc.gamma.RI, Fc.gamma.RIIa, Fc.gamma.RIIIa and Fc.gamma.RIIIb receptors but maintain binding to human Fc.gamma.RIIb receptor and optionally maintain binding to human FcRn receptor. Fc.gamma.RI, Fc.gamma.RIIa, Fc.gamma.RIIIa and Fc.gamma.RIIIb are examples of activating receptors. Fc.gamma.RIIb is an example of an inhibitory receptor. FcRn is an example of a recycling receptor. In some aspects, binding affinity of an anti-ERBB3 antibody or an antigen-binding portion thereof for human Fc receptors may be measured by BIACORE.RTM. analysis. In some aspects, Homogeneous Time Resolved Fluorescence (HTRF) can be used to study binding of an anti-ERBB3 antibody to human Fc receptors. In one example of HTRF, human IgG1 (wild type) is labelled, as is the full suite of Fc gamma receptors and then antibodies with engineered Fc fragments are used in titration competition. In some aspects, ERBB3-positive cells may be mixed with human white blood cells and anti-ERBB3 antibodies, and cell killing by CDC, ADCC and/or ADCP may be measured. In some aspects, an anti-ERBB3 antibody comprising an amino acid sequence of an Fc region of human IgG1-3M (see Table 9) is effector null. In some aspects, an anti-ERBB3 antibody comprising an amino acid sequence of an Fc region of human IgG1-3M (see Table 9) is not effector null.
[0206] The antibody molecule or antigen-binding portion thereof may be a Fab fragment, a F(ab)2 fragment, an Fv fragment, a tetrameric antibody, a tetravalent antibody, a multispecific antibody (for example, a bispecific antibody), a domain-specific antibody, a single domain antibody, a monoclonal antibody or a fusion protein. In one embodiment, an antibody may be a bispecific antibody that binds specifically to a first antigen and a second antigen, wherein the first antigen is ERBB3 and the second antigen is not ERBB3. Antibody molecules and methods for their construction and use are described, in for example Holliger & Hudson (2005, Nature Biotechnol. 23(9): 1126-1136).
[0207] In another aspect of the invention, there is provided an immunoconjugate comprising the antibody molecule or antigen-binding portion thereof of the invention as defined herein linked to a therapeutic agent.
[0208] Examples of suitable therapeutic agents include cytotoxins, radioisotopes, chemotherapeutic agents, immunomodulatory agents, anti-angiogenic agents, antiproliferative agents, pro-apoptotic agents, and cytostatic and cytolytic enzymes (for example RNAses). Further therapeutic agents include a therapeutic nucleic acid, such as a gene encoding an immunomodulatory agent, an anti-angiogenic agent, an anti-proliferative agent, or a pro-apoptotic agent. These drug descriptors are not mutually exclusive, and thus a therapeutic agent may be described using one or more of the above terms.
[0209] Examples of suitable therapeutic agents for use in immunoconjugates include the taxanes, maytansines, CC-1065 and the duocarmycins, the calicheamicins and other enediynes, and the auristatins. Other examples include the anti-folates, vinca alkaloids, and the anthracyclines. Plant toxins, other bioactive proteins, enzymes (i.e., ADEPT), radioisotopes, photosensitizers may also be used in immunoconjugates. In addition, conjugates can be made using secondary carriers as the cytotoxic agent, such as liposomes or polymers, Suitable cytotoxins include an agent that inhibits or prevents the function of cells and/or results in destruction of cells. Representative cytotoxins include antibiotics, inhibitors of tubulin polymerization, alkylating agents that bind to and disrupt DNA, and agents that disrupt protein synthesis or the function of essential cellular proteins such as protein kinases, phosphatases, topoisomerases, enzymes, and cyclins.
[0210] Representative cytotoxins include, but are not limited to, doxorubicin, daunorubicin, idarubicin, aclarubicin, zorubicin, mitoxantrone, epirubicin, carubicin, nogalamycin, menogaril, pitarubicin, valrubicin, cytarabine, gemcitabine, trifluridine, ancitabine, enocitabine, azacitidine, doxifluhdine, pentostatin, broxuhdine, capecitabine, cladhbine, decitabine, floxuhdine, fludarabine, gougerotin, puromycin, tegafur, tiazofuhn, adhamycin, cisplatin, carboplatin, cyclophosphamide, dacarbazine, vinblastine, vincristine, mitoxantrone, bleomycin, mechlorethamine, prednisone, procarbazine, methotrexate, flurouracils, etoposide, taxol, taxol analogs, platins such as cis-platin and carbo-platin, mitomycin, thiotepa, taxanes, vincristine, daunorubicin, epirubicin, actinomycin, authramycin, azaserines, bleomycins, tamoxifen, idarubicin, dolastatins/auristatins, hemiasterlins, esperamicins and maytansinoids.
[0211] Suitable immunomodulatory agents include anti-hormones that block hormone action on tumors and immunosuppressive agents that suppress cytokine production, down-regulate self-antigen expression, or mask MHC antigens.
[0212] Also provided is a nucleic acid molecule encoding the antibody molecule or antigen-binding portion thereof of the invention as defined herein. A nucleic acid molecule may encode (a) the VH region amino acid sequence; (b) the VL region amino acid sequence; or (c) both the VH and the VL region amino acid sequences of an anti-ERBB3 antibody or an antigen-binding portion thereof described herein. In some aspects, the nucleic acid molecule as defined herein may be isolated.
[0213] Further provided is a vector comprising the nucleic acid molecule of the invention as defined herein. The vector may be an expression vector.
[0214] Also provided is a host cell comprising the nucleic acid molecule or the vector of the invention as defined herein. The host cell may be a recombinant host cell.
[0215] In a further aspect there is provided a method of producing an anti-ERBB3 antibody and/or an antigen-binding portion thereof, comprising culturing the host cell of the invention under conditions that result in expression and/or production of the antibody and/or the antigen-binding portion thereof, and isolating the antibody and/or the antigen-binding portion thereof from the host cell or culture.
[0216] In another aspect of the invention there is provided a pharmaceutical composition comprising the antibody molecule or antigen-binding portion thereof of the invention as defined herein, or the nucleic acid molecule of the invention as defined herein, or the vector of the invention as defined herein.
[0217] The invention also provides a method for inhibiting ERBB3 signaling in a cell, the method comprising contacting the cell with an anti-ERBB3 antibody molecule or antigen-binding portion thereof described herein. In some embodiments, an anti-ERBB3 antibody molecule or antigen-binding portion of the invention locks ERBB3 into a monomeric form.
[0218] Further provided is a method for enhancing an immune response in a subject, comprising administering to the subject an effective amount of the antibody molecule or antigen-binding portion thereof of the invention as defined herein, or the immunoconjugate of the invention as defined herein, or the nucleic acid molecule of the invention as defined herein, or the vector of the invention as defined herein, or the pharmaceutical composition of the invention as defined herein. In some embodiments, an anti-ERBB3 antibody molecule or antigen-binding portion of the invention engages a subject's immune cells via antibody effector-function mediated engagement.
[0219] In a further aspect there is provided a method for treating or preventing cancer in a subject, comprising administering to the subject an effective amount of the antibody molecule or antigen-binding portion thereof of the invention as defined herein, or the immunoconjugate of the invention as defined herein, or the nucleic acid molecule of the invention as defined herein, or the vector of the invention as defined herein, or the pharmaceutical composition of the invention as defined herein.
[0220] For example, the cancer may be Gastrointestinal Stromal cancer (GIST), pancreatic cancer, melanoma, breast cancer, lung cancer, bronchial cancer, colorectal cancer, prostate cancer, stomach cancer, ovarian cancer, urinary bladder cancer, brain or central nervous system cancer, peripheral nervous system cancer, esophageal cancer, cervical cancer, uterine or endometrial cancer, cancer of the oral cavity or pharynx, liver cancer, kidney cancer, testicular cancer, biliary tract cancer, small bowel or appendix cancer, salivary gland cancer, thyroid gland cancer, adrenal gland cancer, osteosarcoma, chondrosarcoma, or cancer of hematological tissues.
[0221] The invention also provides an antibody molecule or antigen-binding portion thereof of the invention as defined herein, or the immunoconjugate of the invention as defined herein, or the nucleic acid molecule of the invention as defined herein, or the vector of the invention as defined herein, or the pharmaceutical composition of the invention as defined herein, for use in the treatment of cancer.
[0222] In another aspect the invention provides the antibody molecule, or antigen-binding portion thereof, or the immunoconjugate, or the nucleic acid molecule, or the vector for use, or the method of treatment of the invention as defined herein, for separate, sequential or simultaneous use in a combination combined with a second therapeutic agent, for example an anti-cancer agent.
[0223] In a further aspect there is provided the use of an antibody molecule or antigen-binding portion thereof of the invention as defined herein, or an immunoconjugate of the invention as defined herein, or a nucleic acid molecule of the invention as defined herein, or a vector of the invention as defined herein, or a pharmaceutical composition of the invention as defined herein, in the manufacture of a medicament for the treatment of cancer.
[0224] The invention also provides a method for treating or preventing an autoimmune disease or an inflammatory disease in a subject, comprising administering to the subject an effective amount of the antibody molecule or antigen-binding portion thereof as defined herein, or the immunoconjugate as defined here, or the nucleic acid molecule as defined herein, or the vector as defined herein, or the pharmaceutical composition as defined herein.
[0225] For example the autoimmune disease or inflammatory disease may be arthritis, asthma, multiple sclerosis, psoriasis, Crohn's disease, inflammatory bowel disease, lupus, Grave's disease and Hashimoto's thyroiditis, or ankylosing spondylitis.
[0226] Also provided is an antibody molecule or antigen-binding portion thereof as defined herein, or the immunoconjugate as defined herein, or the nucleic acid molecule as defined herein, or the vector as defined herein, or the pharmaceutical composition as defined herein, for use in the treatment of an autoimmune disease or an inflammatory disease.
[0227] Further provided is the use of an antibody molecule or antigen-binding portion thereof as defined herein, or an immunoconjugate as defined herein, or a nucleic acid molecule as defined herein, or a vector as defined herein, or a pharmaceutical composition as defined herein, in the manufacture of a medicament for the treatment of an autoimmune disease or an inflammatory disease.
[0228] The invention also provides a method for treating or preventing a cardiovascular disease or a fibrotic disease in a subject, comprising administering to the subject an effective amount of the antibody molecule or antigen-binding portion thereof as defined herein, or the immunoconjugate as defined here, or the nucleic acid molecule as defined herein, or the vector as defined herein, or the pharmaceutical composition as defined herein.
[0229] Also provided is an antibody molecule or antigen-binding portion thereof as defined herein, or the immunoconjugate as defined herein, or the nucleic acid molecule as defined herein, or the vector as defined herein, or the pharmaceutical composition as defined herein, for use in the treatment of a cardiovascular disease or a fibrotic disease.
[0230] Further provided is the use of an antibody molecule or antigen-binding portion thereof as defined herein, or an immunoconjugate as defined herein, or a nucleic acid molecule as defined herein, or a vector as defined herein, or a pharmaceutical composition as defined herein, in the manufacture of a medicament for the treatment of a cardiovascular disease or a fibrotic disease.
[0231] The cardiovascular disease in any aspect of the invention may for example be coronary heart disease or atherosclerosis.
[0232] For example, the fibrotic disease in any aspect of the invention may be myocardial infarction, angina, osteoarthritis, pulmonary fibrosis, asthma, cystic fibrosis or bronchitis.
[0233] In one embodiment, the invention provides an anti-ERBB3 antibody or an antigen-binding portion thereof comprising the amino acid sequences disclosed herein for use in therapy.
[0234] The pharmaceutical composition of the invention may comprise a pharmaceutically acceptable excipient, carrier or diluent. A pharmaceutically acceptable excipient may be a compound or a combination of compounds entering into a pharmaceutical composition which does not provoke secondary reactions and which allows, for example, facilitation of the administration of the anti-ERBB3 antibody molecule, an increase in its lifespan and/or in its efficacy in the body or an increase in its solubility in solution. These pharmaceutically acceptable vehicles are well known and will be adapted by the person skilled in the art as a function of the mode of administration of the anti-ERBB3 antibody molecule.
[0235] In some embodiments, the anti-ERBB3 antibody molecule may be provided in a lyophilised form for reconstitution prior to administration. For example, lyophilised antibody molecules may be re-constituted in sterile water and mixed with saline prior to administration to an individual.
[0236] The anti-ERBB3 antibody molecules will usually be administered in the form of a pharmaceutical composition, which may comprise at least one component in addition to the antibody molecule. Thus pharmaceutical compositions may comprise, in addition to the anti-ERBB3 antibody molecule, a pharmaceutically acceptable excipient, carrier, buffer, stabilizer or other materials well known to those skilled in the art. Such materials should be non-toxic and should not interfere with the efficacy of the anti-ERBB3 antibody molecule. The precise nature of the carrier or other material will depend on the route of administration, which may be by bolus, infusion, injection or any other suitable route, as discussed below.
[0237] For parenteral, for example sub-cutaneous or intra-venous administration, e.g. by injection, the pharmaceutical composition comprising the anti-ERBB3 antibody molecule may be in the form of a parenterally acceptable aqueous solution which is pyrogen-free and has suitable pH, isotonicity and stability. Those of relevant skill in the art are well able to prepare suitable solutions using, for example, isotonic vehicles, such as Sodium Chloride Injection, Ringer's Injection, Lactated Ringer's Injection. Preservatives, stabilizers, buffers, antioxidants and/or other additives may be employed as required including buffers such as phosphate, citrate and other organic acids; antioxidants, such as ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens, such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3'-pentanol; and m-cresol); low molecular weight polypeptides; proteins, such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagines, histidine, arginine, or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrins; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions, such as sodium; metal complexes (e.g. Zn-protein complexes); and/or non-ionic surfactants, such as TWEEN.TM., PLURONICS.TM. or polyethylene glycol (PEG).
[0238] A pharmaceutical composition comprising an anti-ERBB3 antibody molecule may be administered alone or in combination with other treatments, either simultaneously or sequentially dependent upon the condition to be treated.
[0239] An anti-ERBB3 antibody molecule as described herein may be used in a method of treatment of the human or animal body, including prophylactic or preventative treatment (e.g. treatment before the onset of a condition in an individual to reduce the risk of the condition occurring in the individual; delay its onset; or reduce its severity after onset). The method of treatment may comprise administering the anti-ERBB3 antibody molecule to an individual in need thereof.
[0240] Administration is normally in a "therapeutically effective amount", this being sufficient to show benefit to a patient. Such benefit may be at least amelioration of at least one symptom. The actual amount administered, and rate and time-course of administration, will depend on the nature and severity of what is being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the composition, the method of administration, the scheduling of administration and other factors known to medical practitioners. Prescription of treatment, e.g. decisions on dosage etc., is within the responsibility of general practitioners and other medical doctors and may depend on the severity of the symptoms and/or progression of a disease being treated. Appropriate doses of antibody molecules are well known in the art (Ledermann J. A. et al., 1991, Int. J. Cancer 47: 659-664; Bagshawe K. D. et al., 1991, Antibody, Immunoconjugates and Radiopharmaceuticals 4: 915-922). Specific dosages may be indicated herein or in the Physician's Desk Reference (2003) as appropriate for the type of medicament being administered may be used. A therapeutically effective amount or suitable dose of an antibody molecule may be determined by comparing its in vitro activity and in vivo activity in an animal model. Methods for extrapolation of effective dosages in mice and other test animals to humans are known. The precise dose will depend upon a number of factors, including whether the antibody is for prevention or for treatment, the size and location of the area to be treated, the precise nature of the antibody (e.g. whole antibody, fragment) and the nature of any detectable label or other molecule attached to the antibody.
[0241] A typical antibody dose will be in the range 100 .mu.g to 1 g for systemic applications, and 1 .mu.g to 1 mg for topical applications. An initial higher loading dose, followed by one or more lower doses, may be administered. Typically, the antibody will be a whole antibody, e.g. the IgG1, IgG1null or IgG4 isotype. This is a dose for a single treatment of an adult patient, which may be proportionally adjusted for children and infants, and also adjusted for other antibody formats in proportion to molecular weight. Treatments may be repeated at daily, twice-weekly, weekly or monthly intervals, at the discretion of the physician. The treatment schedule for an individual may be dependent on the pharmocokinetic and pharmacodynamic properties of the antibody composition, the route of administration and the nature of the condition being treated.
[0242] Treatment may be periodic, and the period between administrations may be about two weeks or more, e.g. about three weeks or more, about four weeks or more, about once a month or more, about five weeks or more, or about six weeks or more. For example, treatment may be every two to four weeks or every four to eight weeks. Treatment may be given before, and/or after surgery, and/or may be administered or applied directly at the anatomical site of surgical treatment or invasive procedure. Suitable formulations and routes of administration are described above.
[0243] In some embodiments, anti-ERBB3 antibody molecules as described herein may be administered as sub-cutaneous injections. Sub-cutaneous injections may be administered using an auto-injector, for example for long or short-term prophylaxis/treatment.
[0244] In some embodiments, the therapeutic effect of the anti-ERBB3 antibody molecule may persist for several multiples of the antibody half-life in serum, depending on the dose. For example, the therapeutic effect of a single dose of the anti-ERBB3 antibody molecule may persist in an individual for 1 month or more, 2 months or more, 3 months or more, 4 months or more, 5 months or more, or 6 months or more.
[0245] The invention also provides a method of producing an antibody molecule which specifically binds to human ERBB3 and optionally also to rhesus monkey ERBB3 or an antigen-binding portion thereof, comprising the steps of:
[0246] (1) grafting anti-ERBB3 CDRs from a non-human source into a human v-domain framework to produce a humanized anti-ERBB3 antibody molecule or antigen-binding portion thereof;
[0247] (2) generating a phage library of clones of the humanized anti-ERBB3 antibody molecule or antigen-binding portion thereof comprising one or more mutations in the CDRs;
[0248] (3) selecting the phage library for binding to human ERBB3 and optionally also to rhesus monkey ERBB3;
[0249] (4) screening clones from the selection step (3) having binding specificity to human ERBB3 and optionally also to rhesus monkey ERBB3; and
[0250] (5) producing an antibody molecule which specifically binds to human ERBB3 and optionally also to rhesus monkey ERBB3, or an antigen-binding portion thereof from clones selected from step (4).
[0251] The method may comprise a further step of producing additional clones based on the clones selected in step (4), for example based on further exploratory mutagenesis at specific positions in the CDRs of the clones selected in step (4), to enhance humanization and/or minimise human T cell epitope content and/or improve manufacturing properties in the antibody molecule or antigen-binding portion thereof produced in step (5).
[0252] Refinements applicable to the above method are as described in Example 1 below.
[0253] As used herein, the term "ERBB3" refers to the ERBB3 protein and variants thereof that retain at least part of the biological activity of ERBB3. As used herein, ERBB3 includes all mammalian species of native sequence ERBB3, including human, rat, mouse and chicken. The term "ERBB3" is used to include variants, isoforms and species homologs of human ERBB3. Antibodies of the invention may cross-react with ERBB3 from species other than human, in particular ERBB3 from rhesus monkey (Macaca mulatta). Examples of human and rhesus ERBB3 amino acid sequences are provided in Table 10. In certain embodiments, the antibodies may be completely specific for human ERBB3 and may not exhibit non-human cross-reactivity.
[0254] As used herein, an "antagonist" as used in the context of the antibody of the invention or an "anti-ERBB3 antagonist antibody" (interchangeably termed "anti-ERBB3 antibody") refers to an antibody which is able to bind to ERBB3 and inhibit ERBB3 biological activity and/or downstream pathway(s) mediated by ERBB3 signaling. An anti-ERBB3 antagonist antibody encompasses antibodies that can block, antagonize, suppress or reduce (including significantly) ERBB3 biological activity, including downstream pathways mediated by ERBB3 signaling, such as receptor binding and/or elicitation of a cellular response to ERBB3. For the purposes of the present invention, it will be explicitly understood that the term "anti-ERBB3 antagonist antibody" encompass all the terms, titles, and functional states and characteristics whereby ERBB3 itself, and ERBB3 biological activity, or the consequences of the activity or biological activity, are substantially nullified, decreased, or neutralized in any meaningful degree.
[0255] The antibody "specifically binds" "specifically interacts", "preferentially binds", "binds" or "interacts" with ERBB3 if it binds with greater affinity, avidity, more readily and/or with greater duration than it binds to other receptors.
[0256] An "antibody molecule" is an immunoglobulin molecule capable of specific binding to a target, such as a carbohydrate, polynucleotide, lipid, polypeptide, etc., through at least one antigen recognition site, located in the variable region of the immunoglobulin molecule. As used herein, the term "antibody molecule" encompasses not only intact polyclonal or monoclonal antibodies, but also any antigen binding fragment (for example, an "antigen-binding portion") or single chain thereof, fusion proteins comprising an antibody, and any other modified configuration of the immunoglobulin molecule that comprises an antigen recognition site including, for example without limitation, scFv, single domain antibodies (for example, shark and camelid antibodies), maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, v-NAR and bis-scFv.
[0257] An "antibody molecule" encompasses an antibody of any class, such as IgG, IgA, or IgM (or sub-class thereof), and the antibody need not be of any particular class. Depending on the antibody amino acid sequence of the constant region of its heavy chains, immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), for example IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The heavy-chain constant regions that correspond to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known.
[0258] The term "antigen binding portion" of an antibody molecule, as used herein, refers to one or more fragments of an intact antibody that retain the ability to specifically bind to ERBB3. Antigen binding functions of an antibody molecule can be performed by fragments of an intact antibody. Examples of binding fragments encompassed within the term "antigen binding portion" of an antibody molecule include Fab; Fab'; F(ab')2; an Fd fragment consisting of the VH and CH1 domains; an Fv fragment consisting of the VL and VH domains of a single arm of an antibody; a single domain antibody (dAb) fragment, and an isolated complementarity determining region (CDR).
[0259] The term "Fc region" is used to define a C-terminal region of an immunoglobulin heavy chain. The "Fc region" may be a native sequence Fc region or a variant Fc region. Although the boundaries of the Fc region of an immunoglobulin heavy chain might vary, the human IgG heavy chain Fc region is usually defined to stretch from an amino acid residue at position Cys226, or from Pro230, to the carboxyl-terminus thereof. The numbering of the residues in the Fc region is that of the EU index as in Kabat. The Fc region of an immunoglobulin generally comprises two constant domains, CH2 and CH3. As is known in the art, an Fc region can be present in dimer or monomeric form.
[0260] A "variable region" of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy chain, either alone or in combination. As known in the art, the variable regions of the heavy and light chain each consist of four framework regions (FRs) connected by three complementarity determining regions (CDRs) also known as hypervariable regions, and contribute to the formation of the antigen binding site of antibodies. When choosing FR to flank CDRs, for example when humanizing or optimizing an antibody, FRs from antibodies which contain CDR sequences in the same canonical class are preferred.
[0261] The CDR definitions used in the present application combine the domains used in the many disparate, often conflicting schemes that have been created in the field, which are based on the combination of immunoglobulin repertoire analyses and structural analyses of antibodies in isolation and in their co-crystals with antigens (see review by Swindells et al., 2016, abYsis: Integrated Antibody Sequence and Structure-Management, Analysis, and Prediction. J Mol Biol. [PMID: 27561707; Epub 22 Aug. 2016]). The CDR definition used herein (a "Unified" definition) incorporates the lessons of all such prior insights and includes all appropriate loop positions required to sample the full residue landscape that potentially mediates target-binding complementarity.
[0262] Table 1 shows the amino acid sequences of the 24C05 murine anti-ERBB3 antibody CDRs as defined herein (a "Unified" scheme), in comparison to well-known alternative systems for defining the same CDRs.
[0263] As used herein the term "conservative substitution" refers to replacement of an amino acid with another amino acid which does not significantly deleteriously change the functional activity. A preferred example of a "conservative substitution" is the replacement of one amino acid with another amino acid which has a value.gtoreq.0 in the following BLOSUM 62 substitution matrix (see Henikoff & Henikoff, 1992, PNAS 89: 10915-10919):
TABLE-US-00001 A R N D C Q E G H I L K M F P S T W Y V A 4 -1 -2 -2 0 -1 -1 0 -2 -1 -1 -1 -1 -2 -1 1 0 -3 -2 0 R -1 5 0 -2 -3 1 0 -2 0 -3 -2 2 -1 -3 -2 -1 -1 -3 -2 -3 N -2 0 6 1 -3 0 0 0 1 -3 -3 0 -2 -3 -2 1 0 -4 -2 -3 D -2 -2 1 6 -3 0 2 -1 -1 -3 -4 -1 -3 -3 -1 0 -1 -4 -3 -3 C 0 -3 -3 -3 9 -3 -4 -3 -3 -1 -1 -3 -1 -2 -3 -1 -1 -2 -2 -1 Q -1 1 0 0 -3 5 2 -2 0 -3 -2 1 0 -3 -1 0 -1 -2 -1 -2 E -1 0 0 2 -4 2 5 -2 0 -3 -3 1 -2 -3 -1 0 -1 -3 -2 -2 G 0 -2 0 -1 -3 -2 -2 6 -2 -4 -4 -2 -3 -3 -2 0 -2 -2 -3 -3 H -2 0 1 -1 -3 0 0 -2 8 -3 -3 -1 -2 -1 -2 -1 -2 -2 2 -3 I -1 -3 -3 -3 -1 -3 -3 -4 -3 4 2 -3 1 0 -3 -2 -1 -3 -1 3 L -1 -2 -3 -4 -1 -2 -3 -4 -3 2 4 -2 2 0 -3 -2 -1 -2 -1 1 K -1 2 0 -1 -3 1 1 -2 -1 -3 -2 5 -1 -3 -1 0 -1 -3 -2 -2 M -1 -1 -2 -3 -1 0 -2 -3 -2 1 2 -1 5 0 -2 -1 -1 -1 -1 1 F -2 -3 -3 -3 -2 -3 -3 -3 -1 0 0 -3 0 6 -4 -2 -2 1 3 -1 P -1 -2 -2 -1 -3 -1 -1 -2 -2 -3 -3 -1 -2 -4 7 -1 -1 -4 -3 -2 S 1 -1 1 0 -1 0 0 0 -1 -2 -2 0 -1 -2 -1 4 1 -3 -2 -2 T 0 -1 0 -1 -1 -1 -1 -2 -2 -1 -1 -1 -1 -2 -1 1 5 -2 -2 0 W -3 -3 -4 -4 -2 -2 -3 -2 -2 -3 -2 -3 -1 1 -4 -3 -2 11 2 -3 Y -2 -2 -2 -3 -2 -1 -2 -3 2 -1 -1 -2 -1 3 -3 -2 -2 2 7 -1 V 0 -3 -3 -3 -1 -2 -2 -3 -3 3 1 -2 1 -1 -2 -2 0 -3 -1 4.
[0264] The term "monoclonal antibody" (Mab) refers to an antibody, or antigen-binding portion thereof, that is derived from a single copy or clone, including for example any eukaryotic, prokaryotic, or phage clone, and not the method by which it is produced. Preferably, a monoclonal antibody of the invention exists in a homogeneous or substantially homogeneous population.
[0265] A "humanized" antibody molecule refers to a form of non-human (for example, murine) antibody molecules, or antigen-binding portion thereof, that are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (such as Fv, Fab, Fab', F(ab')2 or other antigen-binding sub-sequences of antibodies) that contain minimal sequence derived from non-human immunoglobulin. Humanized antibodies may be human immunoglobulins (recipient antibody) in which residues from a CDR of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat, or rabbit having the desired specificity, affinity, and capacity.
[0266] "Human antibody or fully human antibody" refers to an antibody molecule, or antigen-binding portion thereof, derived from transgenic mice carrying human antibody genes or from human cells.
[0267] The term "chimeric antibody" is intended to refer to an antibody molecule, or antigen-binding portion thereof, in which the variable region sequences are derived from one species and the constant region sequences are derived from another species, such as an antibody molecule in which the variable region sequences are derived from a mouse antibody and the constant region sequences are derived from a human antibody.
[0268] "Antibody-drug conjugate" and "immunoconjugate" refer to an antibody molecule, or antigen-binding portion thereof, including antibody derivatives that binds to ERBB3, which is conjugated to cytotoxic, cytostatic and/or therapeutic agents.
[0269] Antibody molecules of the invention, or antigen-binding portion thereof, can be produced using techniques well known in the art, for example recombinant technologies, phage display technologies, synthetic technologies or combinations of such technologies or other technologies readily known in the art.
[0270] The term "isolated molecule" (where the molecule is, for example, a polypeptide, a polynucleotide, or an antibody) is a molecule that by virtue of its origin or source of derivation (1) is not associated with naturally associated components that accompany it in its native state, (2) is substantially free of other molecules from the same species (3) is expressed by a cell from a different species, or (4) does not occur 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 also may be rendered substantially free of naturally associated components by isolation, using purification techniques well known in the art. Molecule purity or homogeneity may be assayed by a number of means well known in the art. For example, the purity of a polypeptide sample may be assayed using polyacrylamide gel electrophoresis and staining of the gel to visualize the polypeptide using techniques well known in the art. For certain purposes, higher resolution may be provided by using HPLC or other means well known in the art for purification.
[0271] The term "epitope" refers to that portion of a molecule capable of being recognized by and bound by an antibody molecule, or antigen-binding portion thereof, at one or more of the antibody molecule's antigen-binding regions. Epitopes can consist of defined regions of primary secondary or tertiary protein structure and includes combinations of secondary structural units or structural domains of the target recognised by the antigen binding regions of the antibody, or antigen-binding portion thereof. Epitopes can likewise consist of a defined chemically active surface grouping of molecules such as amino acids or sugar side chains and have specific three-dimensional structural characteristics as well as specific charge characteristics. The term "antigenic epitope" as used herein, is defined as a portion of a polypeptide to which an antibody molecule can specifically bind as determined by any method well known in the art, for example, by conventional immunoassays, antibody competitive binding assays or by x-ray crystallography or related structural determination methods (for example NMR).
[0272] The term "binding affinity" or "KD" refers to the dissociation rate of a particular antigen-antibody interaction. The KD is the ratio of the rate of dissociation, also called the "off-rate (k.sub.off)", to the association rate, or "on-rate (k.sub.on)". Thus, K.sub.D equals k.sub.off/k.sub.on and is expressed as a molar concentration (M). It follows that the smaller the K.sub.D, the stronger the affinity of binding. Therefore, a K.sub.D of 1 .mu.M indicates weak binding affinity compared to a K.sub.D of 1 nM. KD values for antibodies can be determined using methods well established in the art. One method for determining the KD of an antibody is by using surface plasmon resonance (SPR), typically using a biosensor system such as a Biacore.RTM. system.
[0273] The term "potency" is a measurement of biological activity and may be designated as IC.sub.50, or effective concentration of an antibody or antibody drug conjugate to the antigen ERBB3 to inhibit 50% of activity measured in a ERBB3 activity assay as described herein.
[0274] The phrase "effective amount" or "therapeutically effective amount" as used herein refers to an amount necessary (at dosages and for periods of time and for the means of administration) to achieve the desired therapeutic result. An effective amount is at least the minimal amount, but less than a toxic amount, of an active agent which is necessary to impart therapeutic benefit to a subject.
[0275] The term "inhibit" or "neutralize" as used herein with respect to bioactivity of an antibody molecule of the invention means the ability of the antibody to substantially antagonize, prohibit, prevent, restrain, slow, disrupt, eliminate, stop, reduce or reverse for example progression or severity of that which is being inhibited including, but not limited to, a biological activity or binding interaction of the antibody molecule to ERBB3.
[0276] A "host cell" includes an individual cell or cell culture that can be or has been a recipient for vector(s) for incorporation of polynucleotide inserts. Host cells include progeny of a single host cell, and the progeny may not necessarily be completely identical (in morphology or in genomic DNA complement) to the original parent cell due to natural, accidental, or deliberate mutation. A host cell includes cells transfected in vivo with a polynucleotide(s) of this invention.
[0277] As used herein, "vector" means a construct, which is capable of delivering, and, preferably, expressing, one or more gene(s) or sequence(s) of interest in a host cell. Examples of vectors include, but are not limited to, viral vectors, naked DNA or RNA expression vectors, plasmid, cosmid or phage vectors, DNA or RNA expression vectors associated with cationic condensing agents, DNA or RNA expression vectors encapsulated in liposomes, and certain eukaryotic cells, such as producer cells.
[0278] The term "treating", as used herein, unless otherwise indicated, means reversing, alleviating, inhibiting the progress of, delaying the progression of, delaying the onset of, or preventing the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition. The term "treatment", as used herein, unless otherwise indicated, refers to the act of treating as defined above. The term "treating" also includes adjuvant and neoadjuvant treatment of a subject. For the avoidance of doubt, reference herein to "treatment" includes reference to curative, palliative and prophylactic treatment. For the avoidance of doubt, references herein to "treatment" also include references to curative, palliative and prophylactic treatment.
[0279] It is understood that wherever embodiments are described herein with the language "comprising," otherwise analogous embodiments described in terms of "consisting of" and/or "consisting essentially of" are also provided.
[0280] Where aspects or embodiments of the invention are described in terms of a Markush group or other grouping of alternatives, the present invention encompasses not only the entire group listed as a whole, but each member of the group individually and all possible subgroups of the main group, but also the main group absent one or more of the group members. The present invention also envisages the explicit exclusion of one or more of any of the group members in the claimed invention.
[0281] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control. Throughout this specification and claims, the word "comprise," or variations such as "comprises" or "comprising" will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers. Unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. Any example(s) following the term "e.g." or "for example" is not meant to be exhaustive or limiting.
[0282] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are within the skill of the art.
[0283] Particular non-limiting embodiments of the present invention will now be described with reference to accompanying drawings.
EXAMPLE 1
Generation of Optimized Anti-ERBB3 Therapeutic Antibodies
[0284] Introduction
[0285] In this example, we successfully generated a panel of antagonistic, optimized anti-ERBB3 antibodies. These anti-ERBB3 antibodies are well expressed, biophysically stable, highly soluble and of maximized amino acid sequence identity to preferred human germlines.
[0286] Materials and Methods
[0287] ERBB3 Library Generation and Selection
[0288] The ERBB3 Fab mutagenesis repertoire was assembled by mass oligo synthesis and PCR. This library was designed to sample the germline human CDR residue or murine CDR residue at all positions where the sequences differed, but also sampled all amino acids other than cysteine at key selected CDR positions e.g. in HCDR1 and HCDR3. The amplified Fab repertoire was then cloned via restriction-ligation into a phagemid vector, transformed into E. coli TG-1 cells, and the phage repertoire rescued essentially as previously described in detail (Finlay et al., 2011, Methods Mol Biol 681: 383-401).
[0289] Phage selections were performed by coating streptavidin magnetic microbeads with biotinylated ERBB3 target protein (either human or rhesus), washing the beads thrice with PBS and resuspending in PBS pH7.4 plus 5% skim milk protein. These beads were coated at 100 nM target protein in round 1 of selection, followed by reduced antigen concentrations in three successive rounds. In each round, phage were eluted using trypsin before re-infection into TG1 cells.
[0290] Periplasmic Extracts Production (Small-Scale)
[0291] Production of soluble Fabs in individual E. coli clones was performed. E. coli TG1 cells in logarhythmic growth phase were induced with isopropyl 1-thio-.beta.-D-galactopyranoside. Periplasmic extracts containing soluble Fab were generated by a freeze/thaw cycle: Bacterial cell pellets were frozen at -20.degree. C. for overnight and then thawed at room temperature and resuspended in PBS pH 7.4. The supernatants containing the soluble Fab were collected after shaking at room temperature and centrifugation.
[0292] IgG Expression and Purification
[0293] Mammalian codon-optimized synthetic genes encoding the heavy and light chain variable domains of the lead panel anti-ERBB3 antibodies plus the m24C05 and h24C05 were cloned into mammalian expression vectors comprising IgG1null (`IgG1null`; human IgG1 containing L234A, L235A, G237A mutations in the lower hinge that abrogate normal immunoglobulin Fc effector functions) or IgG1 and human C.kappa. domains, respectively. Co-transfection of heavy and light chain containing vector in mammalian expression system was performed, followed by protein A-based purification of the IgG, quantification and QC on denaturing and non-denaturing SDS-PAGE.
[0294] Direct Binding ELISA for Fab and IgG
[0295] Binding and cross-reactivity of the lead panel to the recombinant proteins was initially assessed by binding ELISA. The human ERBB3 human Fc tagged recombinant protein and the rhesus monkey ERBB3 human Fc tagged recombinant protein were coated to the surface of MaxiSorp.TM. flat-bottom 96 well plate at 1 .mu.g/ml. The purified IgG samples were titrated in two fold serial dilutions starting from 500 nM to 0.008 nM and allowed to bind to the coated antigens. The Fabs were detected using mouse anti-c-myc antibody followed by donkey anti-mouse IgG conjugated to horseradish peroxidase. The IgGs were detected using the mouse anti-human IgG conjugated to horseradish peroxidase. Binding signals were visualized with 3,3',5,5'-Tetrannethylbenzidine Substrate Solution (TMB) and the absorbance measured at 450 nm.
[0296] Alphascreen Epitope Competition Assay for IgG1nullnull Antibodies
[0297] The AlphaScreen assay (Perkin Elmer) was performed in a 25 .mu.l final volume in 384-well white microtiter plates (Greiner). The reaction buffer contained 1.times. PBS pH 7.3 (Oxoid, Cat. nr. BR0014G) and 0.05% (v/v) Tween.RTM. 20 (Sigma, Cat. nr. P9416). Purified IgG samples were titrated in three fold serial dilutions starting at 100 nM final concentration and incubated with biotinylated human ERBB3-His (Acrobiosystems) at 1 nM final concentration for 20 minutes at room temperature. The parental IgG and the anti-human IgG1 Acceptor beads at were added and the mix was incubated for 1 hour at room temperature. Followed by addition of the Streptavidin Donor beads and incubation for 30 minutes at room temperature. The emission of light was measured in the EnVision multilabel plate reader (Perkin Elmer) and analysed using the EnVision manager software. Values were reported as Counts Per Second (CPS) and corrected for crosstalk.
[0298] Biacore.RTM. Analyses of IgG Affinity for Monomeric Human and Rhesus ERBB3 in Solution
[0299] Affinity (KD) of purified IgGs was determined via SPR with antigen in-solution on a Biacore.RTM. 3000 (GE). A mouse anti-human antibody (CH1 specific) was immobilized on a CM5 Sensor Chip to a level of 2000 RU in acetate buffer at pH 4.5 using amine coupling following the Wizard instructions for two channels. One channel was used for background signal correction. The standard running buffer HBS-EP pH 7.4 was used. Regeneration was performed with a single injection of 10 .mu.l of 10 mM Glycine at pH 1.5 at 20 .mu.l/minute. IgG samples were injected for 2 minutes at 50 nM at 30 .mu.l/min followed by an off-rate of 60 seconds. The monomeric antigen (human ERBB3 His tagged or rhesus monkey ERBB3 His tag) was injected in two fold serial dilutions from 3 nM down to 0.2 nM, for 2 minutes at 30 .mu.l/min followed by an off-rate of 300 seconds. The obtained sensorgrams were analysed using the Biacore.RTM. 3000 evaluation (BIAevaluation) software. The KD was calculated by simultaneous fitting of the association and dissociation phases to a 1:1 Langmuir binding model.
[0300] Flow Cytometry of IgGs
[0301] Purified IgGs were tested in FACs for binding to human and rhesus monkey ERBB3 expressed on transiently transfected HEK-293 cells and HEK-293 wild-type cells. The IgG samples were titrated in three-fold serial dilutions starting at 500 nM to 0.008 nM. Binding of IgGs was detected with a mouse anti-human IgG conjugated to FITC. Results were analyzed by examining the Mean Fluorescence Intensity (MFI) of 10000 cells per sample in the BL-1 channel detector of a flow cytometer (Attune.TM. NxT Acoustic Focusing Cytometer, Invitrogen/ThermoFisher Scientific). The EC50 values were calculated using the MFI values in GraphPad Prism software (GraphPad Software, La Jolla, Calif.) and 4 parameters.
[0302] Antibody v-Domain T Cell Epitope Content: In Silico Analyses
[0303] In silico technologies (Abzena, Ltd.), which are based on identifying the location of T cell epitopes in therapeutic antibodies and proteins, were used for assessing potential immunogenicity in antibody v-domains. iTope.TM. was used to analyse the VL and a VH sequences of key leads for peptides with promiscuous high affinity binding to human MHC class II. Promiscuous high affinity MHC class II binding peptides are thought to correlate with the presence of T cell epitopes that are high risk indicators for clinical immunogenicity of drug proteins. The iTope.TM. software predicts favourable interactions between amino acid side chains of a peptide and specific binding pockets (in particular pocket positions; p1, p4, p6, p7 and p9) within the open-ended binding grooves of 34 human MHC class II alleles. These alleles represent the most common HLA-DR alleles found world-wide with no weighting attributed to those found most prevalently in any particular ethnic population. Twenty of the alleles contain the `open` p1 configuration and 14 contain the `closed` configuration where glycine at position 83 is replaced by a valine. The location of key binding residues is achieved by the in silico generation of 9mer peptides that overlap by eight amino acids spanning the test protein sequence. This process successfully discriminates with high accuracy between peptides that either bind or do not bind MHC class II molecules.
[0304] In addition, the sequences were analysed using TCED.TM. (T Cell Epitope Database.TM.) search for matches to T cell epitopes previously identified by in vitro human T cell epitope mapping analyses of other protein sequences. The TCED.TM. is used to search any test sequence against a large (>10,000 peptides) database of peptides derived from unrelated protein and antibody sequences.
[0305] Results and Discussion
[0306] CDR Grafting Onto Preferred Human Germline v-Genes
[0307] The CDRs of an antagonistic murine anti-ERBB3 IgG 24C05 (24C05; see WO2011136911A2 and Table 2) were initially introduced to human germline immunoglobulin v-domain framework sequence scaffolds using CDR grafting. To bias our engineering efforts towards final lead therapeutic IgG compounds with optimal drug-like properties, we chose to graft the CDRs of the parental antibody onto "preferred" germline scaffolds IGHV3-11 and IGKV1-39, which are known to have good solubility, high physical stability and are used at high frequency in the expressed human antibody repertoire.
[0308] Those scaffolds and grafted CDR definitions are outlined in Table 2. While this process of CDR grafting is well known, it is still problematic to predict whether a given set of human v-domain sequences will act as suitable acceptor frameworks for non-human CDR grafting. The use of unsuitable frameworks can lead to the loss of target binding function, protein stability issues or even impaired expression of the final IgG. The IGHV3-11/IGKV1-39 graft was therefore taken forward as the template for CDR mutagenesis and selection of improved clones.
[0309] Library Generation and Screening
[0310] The CDR-grafted IGKV1-39/IGHV3-11 v-domain sequences were combined into a Fab phage display format and a mutagenesis library cassette was generated by oligo synthesis and assembly. The final Fab library was ligated into a phage display vector and transformed into E. coli via electroporation to generate 1.2.times.10.sup.9 independent clones. Library build quality was verified by sequencing 96 clones, across both v-domains. This sequencing data showed that the positions encoding either the murine or human germline residue at each position of variance had been effectively sampled at a frequency of approximately 50% and that positions intended to encode all amino acids exhibited full coverage. Libraries were rescued using helper phage M13 and selections performed on biotinylated human and rhesus monkey ERBB3-Fc proteins in multiple separate branches.
[0311] Post-selection screening and DNA sequencing revealed the presence of 658 human and rhesus ERBB3-binding Fab clones that exhibited strong binding to human and rhesus ERBB3 in ELISA (FIG. 1A) and >50% inhibition of 24C05 IgG1null binding to human ERBB3 in Alphascreen assay (FIG. 1B). Amongst these 658 clones, the framework sequences remained fully germline while humanizing mutations were also observed in all CDRs (Table 3). Lead clones were ranked based on level of CDR germlining versus ELISA and Alphascreen signals for binding to both human and rhesus ERBB3-Fc. The v-domains of the 9 top clones from this ranking were then sub-cloned into IgG expression vectors for further testing as below (Table 4).
[0312] While germ-lining mutations were observed in all CDRs for the lead clones derived directly from library selections, it remained possible that sequence analyses might allow further clones to be designed to have maximal humanization. The 658 sequence-unique hits with binding signals against human and rhesus protein were therefore used to analyse the retention frequency for murine amino acids in the CDRs of this functionally characterized population. Positional amino acid retention frequency was expressed as a percentage found in the VL and VH domains (FIGS. 2A&B, respectively). Murine residues with RF<75% were regarded as positions that are possibly not essential to the target-binding paratope and are likely to be open to germ-lining, in a series of combinatorial designs (Table 4). In a surprising finding for such a high-affinity starting clone, only a minority of murine residues were found to be highly positively selected. Indeed, only 3 of the 9 murine residues in the HCDR1 and HCDR2 exhibited retention frequency above 75% (FIG. 2A). This analysis strongly suggested that the VH sequence outside the HCDR3 could possibly be rendered very close in germline identity to IGHV3-11. In the VL domain, only 4 of 10 murine CDR residues derived from the h24C05 sequence were retained with frequencies>75% (FIG. 3A).
[0313] Designs containing combinations of those murine residues with RF>75% with those also heavily-selected in the lead clone population were given the prefix "MH" (MH=Maximally Humanized). In total 5 MH clones were generated. In addition, a `TTP` (TTP=Total Theoretically Possible) clone was generated which combined the 6 most humanized CDR sequences found in the high-functioning, epitope-competing Fab screen. The MH, TTP and library-derived clone v-domains (Table 4) were generated by gene synthesis and (along with the control antibodies), cloned into human expression vectors for production in IgG1null format. All IgGs were readily expressed and purified from transient transfections of mammalian cells.
[0314] Lead IgG Specificity and Potency Characteristics
[0315] The purified IgGs described above were then tested for binding to human and rhesus ERBB3 in direct titration ELISA format (FIGS. 3A&B). This analysis demonstrated that the majority of library derived and designer (MH) clones retained binding activity for human and/or rhesus ERBB3 that was equivalent to, or improved over, the h24C05 IgG1null.
[0316] An Alphascreen assay was established to allow the testing of IgGs for epitope competition with h24C05 IgG binding to biotinylated monomeric human ERBB3. In this assay, the top-performing library-derived and designer IgGs were more effectively differentiated. While all clones exhibited full, concentration-dependent neutralisation, and the majority of clones exhibited equivalent or improved competition for the h24C05 epitope over h24C05 (FIG. 4), some exhibited less potent epitope competition including MH1.
[0317] Biacore.RTM. analyses of binding affinity were performed for all IgGs to solution-phase, monomeric human and rhesus ERBB3 proteins. In all cases, accurate 1:1 binding affinities with low Chi.sup.2 values were obtained (Table 5). These analyses showed that library-derived clones which consistently gave the highest EC50 and IC50 values in Fab and IgG ELISA and Alphascreen assays also showed highest affinity binding to human and rhesus ERBB3 (Table 5). Importantly, these improvements in affinity were recapitulated in rhesus binding, with the majority of these clones exhibiting affinities within 3-fold of the human ERBB3 affinity. Affinity differentials of less than 4-fold between human and rhesus target orthologs are highly beneficial in pre-clinical drug development analyses as they allow significantly better design and interpretation of e.g. monkey safety, PK and PD modelling experiments.
[0318] In addition, comparison of the affinities of MH and TTP clones confirmed the influence of the LCDR1 in maintaining binding affinity, as mutations of the residues `S>N` at positions 11 in that CDR resulted in approximately 2-fold loss of KD for clones TTP in comparison to clone MH5, against human ERBB3 (Table 5). Comparison of clones MH2 and MH3 also confirmed that the mutation of HCDR1 residue 8 (A to S) and HCDR2 residue 10 (Y to T) in MH3 also led to an approximately 3-fold reduction in binding affinity for human and rhesus ERBB3 in comparison to clone MH2 (Table 5).
[0319] The findings outlined above confirmed that the multiple clones could retain high binding affinity (in the pM range), epitope specificity and species cross-reactivity of h24C05, while retaining only minimal non-germline amino acid content in the VH and VL domains (excluding the HCDR3, for which there is no corresponding germline). This near-complete germlining of the VH domain significantly reduced immunogenic potential in man. In addition, the gemlining mutations observed in multiple clones led to the removal or improvement of several amino acid degradation motifs found in the murine CDRs of h24C05 that are a known risk for manufacturing and clinical development qualities in antibody molecules: A putative isomerization risk at LCDR2 position 6 (D) was removed via mutation to Q, a `DG` aspartic acid isomerisation motif in HCDR2 position 6 and 7 was converted to the lower-risk motif `DS`, and a putative oxidation risk at HCDR3 position 2 (W) was removed by mutation to Y or L. These improvements in primary sequence were not possible to predict a priori and are of direct consequence in both manufacturing and clinical development of an antibody therapeutic as they are all potential stability risk motifs, leading to intrinsic product heterogeneity. Such risk motifs can lead to costly development issues where multiple process modifications must be made to maximise intact antibody yield and to minimise product heterogeneity. Degradation motifs are also a clinical development risk, as accelerated antibody breakdown in the body can reduce both half-life and potency of the molecule.
[0320] Flow Cytometric Analyses of Lead IgG Binding Specificity at the Cell Membrane
[0321] Antibodies to ERBB3 were analysed for concentration-dependent binding at the cell surface via flow cytometry. HEK-293 cells were transiently transfected with either human or rhesus ERBB3 full-length cDNAs. Anti-ERBB3 IgGs and an isotype control were then all tested in IgG1null format, over a concentration range of 500-0.008 nM for binding to human (FIG. 5A) and rhesus (FIG. 5B) transfected cells. All IgGs other than the isotype control showed strong concentration-dependent binding to human and rhesus ERBB3+ cells, with a maximum MFI in each case being >20-fold higher than observed background signals for IgG1 isotype control.
[0322] Antibody v-Domain T Cell Epitope Analyses
[0323] In silico technologies (Abzena, Ltd.), which are based on identifying the location of T cell epitopes in therapeutic antibodies and proteins, were used for assessing the immunogenicity of both the h24C05 and lead antibody 15G11 v-domains. Analysis of the v-domain sequences was performed with overlapping 9mer peptides (with each overlapping the last peptide by 8 residues) which were tested against each of the 34 MHC class II allotypes. Each 9mer was scored based on the potential `fit` and interactions with the MHC class II molecules. The peptide scores calculated by the software lie between 0 and 1. Peptides that produced a high mean binding score (>0.55 in the iTope.TM. scoring function) were highlighted and, if >50% of the MHC class II binding peptides (i.e. 17 out of 34 alleles) had a high binding affinity (score>0.6), such peptides were defined as `high affinity` MHC class II binding peptides which are considered a high risk for containing CD4+ T cell epitopes. Low affinity MHC class II binding peptides bind a high number of alleles (>50%) with a binding score>0.55 (but without a majority >0.6). Further analysis of the sequences was performed using the TCED.TM.. The sequences were used to interrogate the TCED.TM. by BLAST search in order to identify any high sequence homology between peptides (T cell epitopes) from unrelated proteins/antibodies that stimulated T cell responses in previous in vitro T cell epitope mapping studies performed at Abzena Ltd.
[0324] Peptides were grouped into four classes: High Affinity Foreign (`HAF`--high immunogenicity risk), Low Affinity Foreign (`LAF`--lower immunogenicity risk), TCED+ (previously identified epitope in TCED database), and Germline Epitope (`GE`--human germline peptide sequence with high MHC Class II binding affinity). Germline Epitope 9mer peptides are unlikely to have immunogenic potential due to T cell tolerance, as validated by previous studies with a wide range of germline peptides. Importantly, such germline v-domain epitopes (aided further by similar sequences in the human antibody constant regions) also compete for MHC Class II occupancy at the membrane of antigen presenting cells, reducing the risk of foreign peptide presentation being sufficient to achieve the `activation threshold` required for T cell stimulation. High GE content is therefore a beneficial quality in clinical development of an antibody therapeutic.
[0325] This analysis showed that despite the replacement with several murine residues with human germline equivalents in the CDRs of the key lead 15G11, the method of Townsend et al. had not led to significant beneficial changes in peptide epitope content in comparison to h24C05 (FIG. 6A, FIG. 6B). On the contrary, while 15G11 had improved GE content (11) over h24C05 (10), 15G11 had unexpectedly increased immunogenicity risk rather than decreased, as it contained not only 2 new LAF epitopes, but had also gained a high-risk TCED+ LAF in the HCDR-1. As the v-domain framework regions (i.e. outside the CDR sequences) of both antibodies were germline in sequence (Table 2), all changes in predicted immunogenicity in 15G11 came about as a result of the germlining of CDR residues.
[0326] As the method of Townsend et al. had failed to improve either the potency or immunogenicity risk of lead clone 15G11 over h24C05, the potential of further non-human germline mutagenesis in the CDRs was examined. Selection of specific amino acid changes was influenced by the available biophysical and biochemical data, e.g. constraints on modification of the parental 15G11 sequence taking into consideration secondary and tertiary protein structures as well as potential interactions of amino acid side chains with the core of the protein. Additionally, selection of amino acid changes was influenced by the frequency of occurrence of any particular amino acid at any given position in the human repertoire. The aim was therefore to avoid introducing amino acids that never occur at a given position and that would be more likely to adversely affect the structure. The amino acid cysteine was not considered at any point to avoid introducing an unpaired cysteine residue, which could potentially lead to aggregation issues. Each epitope was analysed individually to identify residues that would remove or reduce promiscuous MHC class II binding and, subsequently, the proposed epitope variants were analysed in the context of the whole 15G11 sequence to ensure that novel potential epitopes were not introduced in adjacent regions. Increased value was placed on the use of mutations that had been observed to be tolerated in the functionally-selected population of CDR sequences (Table 3). This process was applied to four key predicted epitopes.
[0327] Epitope 1 was the highest risk (TCED+) epitope identified and partially overlaps with VH CDR1 (FIG. 7A). As such, performing any substitutions within this region could potentially impact on binding to antigen. By iTope.TM. analysis, this region consists of one T cell epitope with a p1 anchor at Y32 (Kabat numbering). Only a limited number of changes were assessed for Framework 2 as this region is highly conserved between antibodies, with several residues playing a role in forming the VH:VL interface. In contrast to Framework regions, CDRs show greater sequence diversity, although this diversity is substantially less pronounced for VH CDR1 and 2 compared with VH CDR3. Within the VH CDR1 region, it was observed that for a number of given positions, several amino acid substitutions were able to completely abrogate potential T cell epitope binding. However, in many cases, these amino acids are almost never observed in that given position, and so these amino acids were discarded. Suggested sequence changes are illustrated in FIG. 7A. Two epitope-ablating variants were prioritised in this peptide: YSMSWIRQA (SED ID NO:250) and YGMSWVRQA (SEQ ID NO:251) (mutation underlined in both cases). The first mutation G>S could potentially ablate the TCED+ epitope, whereas the second mutation I>V could potentially render the peptide sequence a GE.
[0328] Epitope 2 lies primarily within VL CDR1 and, as such, performing any substitutions within this region could potentially impact on binding to antigen. By iTope.TM. analysis, this region consists of one potential T cell epitope with a p1 anchor at I29. Changes were kept to a minimum in Framework 2 as this region is very highly conserved between antibodies, with several residues playing a role in forming the VH:VL interface. In contrast to Framework regions, CDRs show greater sequence diversity, although this diversity is substantially less pronounced for VLs compared with VHs and for VL CDR1 and 2 compared with VL CDR3. Within the VL CDR1 region, it was observed that for a number of given positions, several amino acid substitutions were able to completely abrogate potential T cell epitope binding. However, in many cases, these amino acids are almost never observed in that given position, and so these amino acids were discarded. Suggested sequence changes are illustrated in FIG. 7B.
[0329] Epitope 3 overlaps partially with VL CDR2 and, as such, performing any mutations within this region could potentially impact on binding to antigen. By iTope.TM. analysis, this region consists of one potential T cell epitope with a p1 anchor at I48. Changes to Framework 2 were kept to a minimum as this region is highly conserved between antibodies. In contrast to Framework regions, CDRs show greater sequence diversity, although this diversity is less pronounced for VL CDRs compared with VH CDRs. From iTope.TM. analysis, it was observed that the majority of amino acid substitutions at almost all positions within the core 9-mer were detrimental, with increased binding observed for almost all substitutions. A limited number of amino acid substitutions were identified that abrogate potential T cell epitope binding, however, these amino acids are almost never observed in that given position, and so these amino acids were discarded. Suggested sequence changes are illustrated in FIG. 7C. Importantly, in the analysis, as so many changes in this region were potentially detrimental, selected full CDR variants in the LCDR-2 (Tables 3 and 4) were examined for their TCR epitope risk. Surprisingly, the LCDR-2 sequence AASTLQS (SEQ ID NO:26) was found to fully ablate the HAF peptide risk of this epitope, and a similar high-risk HAF epitope in h24C05 (IYAASTLDS (SEQ ID NO:252)). As a result, this CDR sequence was prioritised for inclusion in a subset of new variants.
[0330] Epitope 4 lies completely within VL CDR3 and, as such, performing any substitutions within this region could potentially impact on binding to antigen. By iTope.TM. analysis, this region consists of one potential T cell epitope with a p1 anchor at L89. No Framework changes were considered. In contrast to Framework regions, CDRs show greater sequence diversity, especially for CDR3, and so amino acid substitutions were not discarded in this region on the basis of occurrence. Several single amino acid substitutions were observed to partially abrogate the binding of both potential T cell epitopes. Suggested single amino acid substitution changes that have an effect on Epitope 4 are illustrated in FIG. 7D.
[0331] As there were a number of potential variants to be sampled in each epitope, 11 new variants of 15G11 were designed in an attempt to find an ideal combination that could retain potency but lead to a fully deimmunised clone. For these 15 IgGs the VL v-domain sequences are shown in Table 6, with the VH domains in Table 7. These IgGs were cloned and expressed in IgG1 form, before functional testing as below.
[0332] 15G11-DI Variants--IgG Specificity and Potency Characteristics
[0333] `DI` variant IgGs 15G11-DI1-11 were first examined for their binding characteristics by ELISA on human (FIG. 8A) and rhesus (FIG. 8B) ErbB3. Against both human and rhesus ErbB3, all clones other than 15G11-DI11 exhibited strong target binding with curves overlapping with both m24C05 and h24C05.
[0334] As ELISA binding is strongly influenced by avidity effects, we then performed a high-sensitivity, solution-phase, Alphascreen competition assay to simultaneously examine the target binding affinity and epitope competition retention with h24C05 (FIG. 9). In this assay, all IgGs other than 15G11-DI11 showed full, concentration dependent inhibition of h24C05 to human Erb63, with potencies generally being equivalent to, or improved over h24C05 (FIG. 9). IgGs m24C05 and h24C05 exhibited IC50 values of 0.16 nM and 1.25 nM, respectively, which correlates with the Biacore.RTM. affinity value differential between those two IgGs shown in Table 5. Exceptions were 15G11-DI3, 15G11-DI4, 15G11-DI10 and 15G11-DI11, whose potencies appeared to be lower than h24C05.
[0335] In flow cytometric analyses of binding to human and rhesus Erb63 (FIG. 10A, B), all clones showed fully saturating, overlapping binding curves. Clones 15G11-DI5, 15G11-DI6, 15G11-DI7, 15G11-DI8 and 15G11-DI9 exhibited highest binding to both human and rhesus Erb63, approximately equivalent to both m24C05 and h24C05.
[0336] A key developability factor in CDR-engineered antibodies for therapeutic use is a lack of `polyreactivity`. Polyreactivity is a significant risk to the pharmacokinetic and safety qualities of any antibody. To ensure the deimmunised lead antibodies were not at risk, they were examined in established polyreactivity binding ELISA assays against dsDNA and human insulin (Avery et al. mAbs, 2018). The majority of IgGs tested in this assay (15G11-DI10 being an exception) showed no sign of polyreactive binding as they exhibited signals below the risk cut-off of 10 and at, or below, the signals of the negative control, clinically successful antibodies Bevacizumab and Ustekinumab (FIG. 11). The positive control antibodies Bococizumab and Briakinumab exhibited the expected strong signals in these assays.
[0337] To ascertain if the full biological potency of Erb63 inhibition had been retained in the key deimmunised leads, clones 15G11, 16B09, 15G11-DI5, 15G11-DI6, 15G11-DI7, 15G11-DI8 and 15G11-DI9 were all analysed over a broad concentration range in the DiscoverX PathHunter eXpress Erb62-Erb63 functional assay (FIG. 12A-G). These analyses demonstrated that clones 15G11 (FIG. 12A), 16B09 (FIG. 12B), 15G11-DI5 (FIG. 12C) and 15G11-DI6 (FIG. 12D) all maintained near-identical potency to both m24C05 and h24C05. Clones 15G11-DI7 (FIG. 12E) and 15G11-DI8 (FIG. 12F) exhibited slightly reduced potency. For 15G11-DI9, the curve fit of its data was unreliable (FIG. 12C), so the experiment was repeated and the fit improved, with 15G11-DI9 also showing near identical potency to h24C05 and m24C05, as evidenced by the overlapping response curves (FIG. 13).
[0338] For these deimmunised key leads, TCR epitope content was again examined (FIG. 14A-FIG. 14E) and this demonstrated that clones 15G11-DI8 and 15G11-DI9 exhibited significantly reduced immunogenicity risk in comparison to h24C05 and 15G11 where the initial germlining process had, unexpectedly, actually increased risk over h24C05 (FIG. 6A, FIG. 6B). Indeed, clone 15G11-DI9 was found to be fully inert, containing no predicted foreign TCR epitopes at all and only a large number of GE peptides (11).
[0339] The combined analyses outlined herein demonstrated that, surprisingly, deep sampling of both germline and non-germline amino acids in the CDRs of these antibodies allowed the simultaneous optimisation of target binding specificity, immunogenicity risk, potency, biophysical stability and chemical stability risks in multiple final molecules. These findings also demonstrate that the beneficial deimmunising outcomes described here could not be achieved using the method of Townsend et al. (2015; PNAS 112: 15354-15359).
[0340] Although the present invention has been described with reference to preferred or exemplary embodiments, those skilled in the art will recognize that various modifications and variations to the same can be accomplished without departing from the spirit and scope of the present invention and that such modifications are clearly contemplated herein. No limitation with respect to the specific embodiments disclosed herein and set forth in the appended claims is intended nor should any be inferred.
[0341] No limitation with respect to the specific embodiments disclosed herein and set forth in the appended claims is intended nor should any be inferred. All documents, or portions of documents, cited herein, including but not limited to patents, patent applications, articles, books, and treatises, are hereby expressly incorporated by reference in their entirety for any purpose. In the event that one or more of the incorporated documents or portions of documents define a term that contradicts that term's definition in the application, the definition that appears in this application controls. However, mention of any reference, article, publication, patent, patent publication, and patent application cited herein is not, and should not be taken as an acknowledgment, or any form of suggestion, that they constitute valid prior art or form part of the common general knowledge in any country in the world.
TABLE-US-00002 TABLE 1 Amino acid sequences of 24C05 anti-ERBB3 CDRs as defined here ("Unified" scheme) in comparison to alternative definitions. Scheme HCDR1 HCDR2 HCDR3 LCDR1 LCDR2 LCDR3 Unified GFTFSDYAMS VSTISDGGTYTYYPDNVKG EWGDYDGFDY RASQEISGYLS AASTLDS LQYDSYPYT (SEQ ID (SEQ ID NO: 65) (SEQ ID NO: 72) (SEQ ID NO: 76) (SEQ ID NO: 80) (SEQ ID NO: 82) NO: 59) Kabat DYAMS TISDGGTYTYYPDNVKG EWGDYDGFDY RASQEISGYLS AASTLDS LQYDSYPYT (SEQ ID (SEQ ID NO: 66) (SEQ ID NO: 72) (SEQ ID NO: 76) (SEQ ID NO: 80) (SEQ ID NO:) NO: 60) Chotia GFTFSDY SDGGTY EWGDYDGFDY RASQEISGYLS AASTLDS LQYDSYPYT (SEQ ID (SEQ ID NO: 67) (SEQ ID NO: 72) (SEQ ID NO: 76) (SEQ ID NO: 80) (SEQ ID NO: 82) NO: 61) IMGT GFTFSDYA ISDGGTYT AREWGDYDGFDY QEISGY AAS LQYDSYPYT (SEQ ID (SEQ ID NO: 68) (SEQ ID NO: 73) (SEQ ID NO: 77) (SEQ ID NO: 82) NO: 62) AHo ASGFTFSDYAMS ISDGGTYTYYPDNVKG EWGDYDGFD ASQEISGY AASTLDS YDSYPY (SEQ ID (SEQ ID NO: 69) (SEQ ID NO: 74) (SEQ ID NO: 78) (SEQ ID NO: 80) (SEQ ID NO: 83) NO: 63) AbM GFTFSDYAMS TISDGGTYTY EWGDYDGFDY RASQEISGYLS AASTLDS LQYDSYPYT (SEQ ID (SEQ ID NO: 70) (SEQ ID NO: 72) (SEQ ID NO: 76) (SEQ ID NO: 80) (SEQ ID NO: 82) NO: 59) Contact SDYAMS VSTISDGGTYTY AREWGDYDGFD ISGYLSWY LLIYAASTLD LQYDSYPY (SEQ ID (SEQ ID NO: 71) (SEQ ID NO: 75) (SEQ ID NO: 79) (SEQ ID NO: 81) (SEQ ID NO: 84) NO: 64)
TABLE-US-00003 TABLE 2 Amino acid sequence of h24C05 anti-ERBB3 v-domains and human germline CDR grafts. Human V DOMAIN germline.sup.1 Amino acid sequence.sup.2 h24C05-VH IGHV3-11 QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYAMSWIRQAPGKGLEWVSTISDGGTYTYY PDNVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREWGDYDGFDYWGQGTLVTVSS (SEQ ID NO: 85) VH graft IGHV3-11.sup.3 QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYAMSWIRQAPGKGLEWVSTISDGGTYTYY PDNVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREWGDYDGFDYWGQGTLVTVSS (SEQ ID NO: 86) h24C05-VL IGKV1-16 DIQMTQSPSSLSASVGDRVTITCRASQEISGYLSWFQQKPGKAPKSLIYAASTLDSGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCLQYDSYPYTFGGGTKVEIK (SEQ ID NO: 87) VL graft IGKV1-39.sup.3 DIQMTQSPSSLSASVGDRVTITCRASQEISGYLSWYQQKPGKAPKLLIYAASTLDSGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCLQYDSYPYTFGGGTKVEIK (SEQ ID NO: 88) .sup.1Human germline definitions used for grafting, based on IMGT system. .sup.2CDR residues are in bold and underlined. As noted above, the "Unified" CDR definitions used in this manuscript are an expanded definition in comparison to the classical Kabat definition. Each sequence above shows the framework regions (FRs) and the CDRs in the following order: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. .sup.3Grafts are fully germline in the framework regions, used as the template for CDR mutant library construction.
TABLE-US-00004 TABLE 3 Amino acid sequences of unique CDRs from 658 unique anti-ERBB3 v-domains. LCDR1 LCDR2 LCDR3 HCDR1 HCDR2 HCDR2 HCDR3 RASQSISSYLS AASSLDS LQYDSTPYT GFTFSDYAMS VGTISDGGTTIYYADNVKG VSTISDSGSYIYYADSVKG EFGDYDGFDF (SEQ ID NO: 16) (SEQ ID NO: 22) (SEQ ID NO: 23) (SEQ ID (SEQ ID NO: 117) (SEQ ID NO: 14) (SEQ ID NO: 109) NO: 181) RASQEISSYLS AASSLQS LQYYSYPYT GFTFSDYEMS VSTISDDGSTTYYADSVKG VSTISDSGSYIYYPDNVKG EFGDYDGFDY (SEQ ID NO: 21) (SEQ ID NO: 17) (SEQ ID NO: 98) (SEQ ID (SEQ ID NO: 118) (SEQ ID NO: 155) (SEQ ID NO: 110) NO: 182) RASQIISSYLS AASTLQS LQYDSTPLT GFTFSDYGMS VSTISDGGSTIYYADNVKG VSTISDSGSYIYYPDSVKG ELGDYDGFDY (SEQ ID NO: 52) (SEQ ID NO: 26) (SEQ ID NO: 18) (SEQ ID (SEQ ID NO: 119) (SEQ ID NO: 156) (SEQ ID NO: 13) NO: 20) RASQSISSYLN AASNLQS QQYDSYPYT GFTFSDYHMS VSTISDGGSTIYYADSVKG VSTISDSGSYTYYADNVKG ELGDYDGWDY (SEQ ID NO: 50) (SEQ ID NO: 93) (SEQ ID NO: 99) (SEQ ID (SEQ ID NO: 120) (SEQ ID NO: 157) (SEQ ID NO: 111) NO: 183) RASQSISGYLN AASSLHS LQYYSTPLT GFTFSDYNMS VSTISDGGSTIYYPDNVKG VSTISDSGSYTYYADSVKG EMGDYDGFDY (SEQ ID NO: 89) (SEQ ID NO: 94) (SEQ ID NO: 53) (SEQ ID (SEQ ID NO: 121) (SEQ ID NO: 54) (SEQ ID NO: 112) NO: 184) RASQEISSYLN EASSLDS LQYDSYPLT GFTFSDYQMS VSTISDGGSTIYYPDSVKG VSTISDSGSYTYYPDNVKG EQGDYDGFDI (SEQ ID NO: 90) (SEQ ID NO: 95) (SEQ ID (SEQ ID (SEQ ID NO: 122) (SEQ ID NO: 158) (SEQ ID NO: 100) NO: 113) NO: 185) RASQSISGYLS AASSLKS QQYDSTPYT GFTFSDYRMS VSTISDGGSTTYYADNVKG VSTISDSGSYTYYPDSVKG EWGDADGFDY (SEQ ID NO: 30) (SEQ ID NO: 96) (SEQ ID (SEQ ID (SEQ ID NO: 123) (SEQ ID NO: 19) (SEQ ID NO: 101) NO: 114) NO: 186) RASQEISGYLN EASSLQS LQSDSTPYT GFTFSDYSMS VSTISDGGSTTYYADSVKG VSTISDSGTTIYYADNVKG EWGDDDGFDY (SEQ ID NO: 91) (SEQ ID NO: 97) (SEQ ID (SEQ ID (SEQ ID NO: 124) (SEQ ID NO: 56) (SEQ ID NO: 102) NO: 24) NO: 187) RASQNISSYLS LQSDSTPLT GFTFSDYTMS VSTISDGGSTTYYPDNVKG VSTISDSGTTIYYADSVKG EWGDEDGFDY (SEQ ID NO: 92) (SEQ ID (SEQ ID (SEQ ID NO: 125) (SEQ ID NO: 159) (SEQ ID NO: 103) NO: 115) NO: 188) QQYDSYPLT GFTFSDYVMS VSTISDGGSTTYYPDSVKG VSTISDSGTTTYYADNVKG EWGDHDGFDY (SEQ ID (SEQ ID (SEQ ID NO: 126) (SEQ ID NO: 160) (SEQ ID NO: 104) NO: 116) NO: 189) LQYYSYPLT VSTISDGGSYIYYADNVKG VSTISDSGTTTYYADSVKG EWGDLDGFDY (SEQ ID (SEQ ID NO: 127) (SEQ ID NO: 161) (SEQ ID NO: 105) NO: 190) LQYYSTPYT VSTISDGGSYIYYADSVKG VSTISDSGTTTYYPDNVKG EWGDMDGFDR (SEQ ID (SEQ ID NO: 128) (SEQ ID NO: 162) (SEQ ID NO: 106) NO: 191) QQYDSTPLT VSTISDGGSYIYYPDNVKG VSTISDSGTTTYYPDSVKG EWGDMDGFDY (SEQ ID (SEQ ID NO: 129) (SEQ ID NO: 163) (SEQ ID NO: 107) NO: 192) LQYDSYHLT VSTISDGGSYIYYPDSVKG VSTISDSGTYIYYADNVKG EWGDNDGFDY (SEQ ID (SEQ ID NO: 130) (SEQ ID NO: 164) (SEQ ID NO: 108) NO: 193) VSTISDGGSYTYYADNVKG VSTISDSGTYIYYADSVKG EWGDQDGFDY (SEQ ID NO: 31) (SEQ ID NO: 165) (SEQ ID NO: 194) VSTISDGGSYTYYADSVKG VSTISDSGTYIYYPDNVKG EWGDSDGFDY (SEQ ID NO: 28) (SEQ ID NO: 166) (SEQ ID NO: 195) VSTISDGGSYTYYPDNVKG VSTISDSGTYIYYPDSVKG EWGDTDGFDY (SEQ ID NO: 131) (SEQ ID NO: 167) (SEQ ID NO: 196) VSTISDGGSYTYYPDSVKG VSTISDSGTYTYYADNVKG EWGDWDGFDY (SEQ ID NO: 57) (SEQ ID NO: 168) (SEQ ID NO: 197) VSTISDGGTTIYYADNVKG VSTISDSGTYTYYADSVKG EWGDYDGCDY (SEQ ID NO: 132) (SEQ ID NO: 169) (SEQ ID NO: 198) VSTISDGGTTIYYADSVKG VSTISDSGTYTYYPDNVKG EWGDYDGFDA (SEQ ID NO: 133) (SEQ ID NO: 170) (SEQ ID NO: 199) VSTISDGGTTIYYPDSVKG VSTISDSGTYTYYPDSVKG EWGDYDGFDD (SEQ ID NO: 134) (SEQ ID NO: 25) (SEQ ID NO: 200) VSTISDGGTTTYYADNVKG VSTISNSGTYTYYADSVKG EWGDYDGFDE (SEQ ID NO: 135) (SEQ ID NO: 171) (SEQ ID NO: 29) VSTISDGGTTTYYADSVKG VSTISSGGSYIYYPDSVKG EWGDYDGFDF (SEQ ID NO: 136) (SEQ ID NO: 172) (SEQ ID NO: 15) VSTISDGGTTTYYPDNVKG VSTISSGGSYTYYPDSVKG EWGDYDGFDH (SEQ ID NO: 137) (SEQ ID NO: 173) (SEQ ID NO: 27) VSTISDGGTTTYYPDSVKG VSTISSSGSYTYYADNVKG EWGDYDGFDI (SEQ ID NO: 138) (SEQ ID NO: 174) (SEQ ID NO: 201) VSTISDGGTYIYYADNVKG VSTISSSGTTTYYADSVKG EWGDYDGFDK (SEQ ID NO: 139) (SEQ ID NO: 175) (SEQ ID NO: 202) VSTISDGGTYIYYADSVKG VSTISSSGTYIYYADNVKG EWGDYDGFDL (SEQ ID NO: 140) (SEQ ID NO: 176) (SEQ ID NO: 203) VSTISDGGTYIYYPDNVKG VSYISDGGSYIYYADSVKG EWGDYDGFDM (SEQ ID NO: 141) (SEQ ID NO: 177) (SEQ ID NO: 204) VSTISDGGTYIYYPDSVKG VSYISDGGSYTYYADNVKG EWGDYDGFDN (SEQ ID NO: 142) (SEQ ID NO: 178) (SEQ ID NO: 55) VSTISDGGTYTYYADNVKG VSYISDGGTTIYYADSVKG EWGDYDGFDQ (SEQ ID NO: 143) (SEQ ID NO: 179) (SEQ ID NO: 205) VSTISDGGTYTYYADSVKG VSYISDSGTYTYYPDSVKG EWGDYDGFDR (SEQ ID NO: 144) (SEQ ID NO: 180) (SEQ ID NO: 206) VSTISDGGTYTYYPDNVKG EWGDYDGFDS (SEQ ID NO: 145) (SEQ ID NO: 207) VSTISDGGTYTYYPDSVKG EWGDYDGFDV (SEQ ID NO: 146) (SEQ ID NO: 208) VSTISDSGSTIYYADNVKG EWGDYDGFDW (SEQ ID NO: 147) (SEQ ID NO: 209) VSTISDSGSTIYYADSVKG EWGDYDGFDY (SEQ ID NO: 58) (SEQ ID NO: 210) VSTISDSGSTIYYPDNVKG EWGDYDGFHY (SEQ ID NO: 148) (SEQ ID NO: 211) VSTISDSGSTIYYPDSVKG EWGDYDGIDY (SEQ ID NO: 149) (SEQ ID NO: 212) VSTISDSGSTTYYADNVKG EWGDYDGLDY (SEQ ID NO: 150) (SEQ ID NO: 213) VSTISDSGSTTYYADSVKG EWGDYDGWDY (SEQ ID NO: 151) (SEQ ID NO: 214) VSTISDSGSTTYYPDNVKG EWGDYDGYDY (SEQ ID NO: 152) (SEQ ID NO: 215) VSTISDSGSTTYYPDSVKG EYGDYDGFDY (SEQ ID NO: 153) (SEQ ID NO: 51) VSTISDSGSYIYYADNVKG MWGDYDGFDY (SEQ ID NO: 154) (SEQ ID NO: 216)
TABLE-US-00005 TABLE 4 Amino acid sequences of CDRs of unique, library-derived and designer, human/rhesus cross-reactive anti-ERBB3 IgGs. Clone LCDR1 LCDR2 LCDR3 HCDR1 HCDR2 HCDR3 15D10 RASQSISSYLS AASTLQS LQYDSTPLT GFTFSDYGMS VSTISDGGSYTYYADNVKG EWGDYDGFDF (SEQ ID NO: 16) (SEQ ID NO: 26) (SEQ ID NO: 18) (SEQ ID NO: 13) (SEQ ID NO: 31) (SEQ ID NO: 15) 17H10 RASQSISGYLS AASTLQS LQYDSTPYT GFTFSDYGMS VSTISDGGSYTYYADSVKG EWGDYDGFDE (SEQ ID NO: 30) (SEQ ID NO: 26) (SEQ ID NO: 23) (SEQ ID NO: 13) (SEQ ID NO: 28) (SEQ ID NO: 29) 09D12 RASQSISSYLN AASSLDS LQYDSTPLT GFTFSDYGMS VSTISDGGSYTYYADSVKG EYGDYDGFDY (SEQ ID NO: 50) (SEQ ID NO: 22) (SEQ ID NO: 18) (SEQ ID NO: 13) (SEQ ID NO: 28) (SEQ ID NO: 51) 15D03 RASQEISSYLS AASSLQS LQYDSTPLT GFTFSDYGMS VSTISDSGSYIYYADSVKG EWGDYDGFDH (SEQ ID NO: 21) (SEQ ID NO: 17) (SEQ ID NO: 18) (SEQ ID NO: 13) (SEQ ID NO: 14) (SEQ ID NO: 27) 11H02 RASQIISSYLS AASSLDS LQYYSTPLT GFTFSDYGMS VSTISDSGSYTYYADSVKG EWGDYDGFDN (SEQ ID NO: 52) (SEQ ID NO: 22) (SEQ ID NO: 53) (SEQ ID NO: 13) (SEQ ID NO: 54) (SEQ ID NO: 55) 15G11 RASQEISSYLS AASSLDS LQYDSTPYT GFTFSDYGMS VSTISDSGSYTYYPDSVKG ELGDYDGFDY (SEQ ID NO: 21) (SEQ ID NO: 22) (SEQ ID NO: 23) (SEQ ID NO: 13) (SEQ ID NO: 19) (SEQ ID NO: 20) 15E02 RASQSISSYLS AASSLQS LQYDSTPLT GFTFSDYGMS VSTISDSGTTIYYADNVKG EYGDYDGFDY (SEQ ID NO: 16) (SEQ ID NO: 17) (SEQ ID NO: 18) (SEQ ID NO: 13) (SEQ ID NO: 56) (SEQ ID NO: 51) 09H02 RASQSISSYLS AASSLQS LQYDSTPLT GFTFSDYSMS VSTISDGGSYTYYPDSVKG ELGDYDGFDY (SEQ ID NO: 16) (SEQ ID NO: 17) (SEQ ID NO: 18) (SEQ ID NO: 24) (SEQ ID NO: 57) (SEQ ID NO: 20) 16B09 RASQEISSYLS AASTLQS LQYDSTPLT GFTFSDYSMS VSTISDSGTYTYYPDSVKG EWGDYDGFDF (SEQ ID NO: 21) (SEQ ID NO: 26) (SEQ ID NO: 18) (SEQ ID NO: 24) (SEQ ID NO: 25) (SEQ ID NO: 15) TTP RASQSISSYLN AASSLQS LQYDSTPLT GFTFSDYGMS VSTISDSGSTIYYADSVKG EYGDYDGFDY (SEQ ID NO: 50) (SEQ ID NO: 17) (SEQ ID NO: 18) (SEQ ID NO: 13) (SEQ ID NO: 58) (SEQ ID NO: 51) MH1 RASQSISSYLS AASSLQS LQYDSTPLT GFTFSDYGMS VSTISDSGSYIYYADSVKG ELGDYDGFDY (SEQ ID NO: 16) (SEQ ID NO: 17) (SEQ ID NO: 18) (SEQ ID NO: 13) (SEQ ID NO: 14) (SEQ ID NO: 20) MH2 RASQSISSYLS AASSLQS LQYDSTPLT GFTFSDYGMS VSTISDSGSYIYYADSVKG EWGDYDGFDF (SEQ ID NO: 16) (SEQ ID NO: 17) (SEQ ID NO: 18) (SEQ ID NO: 13) (SEQ ID NO: 14) (SEQ ID NO: 15) MH3 RASQSISSYLS AASSLQS LQYDSTPLT GFTFSDYSMS VSTISDSGSTIYYADSVKG EWGDYDGFDF (SEQ ID NO: 16) (SEQ ID NO: 17) (SEQ ID NO: 18) (SEQ ID NO: 24) (SEQ ID NO: 58) (SEQ ID NO: 15) MH4 RASQSISSYLS AASSLQS LQYDSTPLT GFTFSDYGMS VSTISDSGSYIYYADSVKG EWGDYDGFDE (SEQ ID NO: 16) (SEQ ID NO: 17) (SEQ ID NO: 18) (SEQ ID NO: 13) (SEQ ID NO: 14) (SEQ ID NO: 29) MH5 RASQSISSYLS AASSLQS LQYDSTPLT GFTFSDYGMS VSTISDSGSTIYYADSVKG EYGDYDGFDY (SEQ ID NO: 16) (SEQ ID NO: 17) (SEQ ID NO: 18) (SEQ ID NO: 13) (SEQ ID NO: 58) (SEQ ID NO: 51)
TABLE-US-00006 TABLE 5 BIACORE .RTM. affinity values for IgG binding to human and rhesus monomeric ERBB3. Human ERBB3 Rhesus ERBB3 Clone name ka (1/Ms) kd (1/s) Chi2 KD (nM) ka (1/Ms) kd (1/s) Chi2 KD (nM) 24C05 4.60E+07 6.70E-04 0.117 0.014 1.80E+07 4.40E-04 0.02 0.024 h24C05 2.50E+07 9.60E-04 0.164 0.039 4.10E+06 7.40E-04 0.037 0.18 15G11 1.60E+07 1.20E-03 0.141 0.078 3.30E+06 1.10E-03 0.041 0.33 16B09 1.60E+07 1.40E-03 0.223 0.089 1.30E+07 2.70E-03 0.044 0.21 15D10 1.40E+07 1.90E-03 0.195 0.14 3.30E+06 1.80E-03 0.057 0.54 17H10 9.30E+06 1.70E-03 0.165 0.19 2.70E+06 1.80E-03 0.035 0.65 15D03 1.10E+07 3.00E-03 0.204 0.26 1.80E+07 4.70E-03 0.117 0.26 MH4 1.10E+07 2.90E-03 0.158 0.26 2.20E+07 4.70E-03 0.099 0.21 MH2 1.10E+07 2.90E-03 0.177 0.27 2.00E+07 4.80E-03 0.099 0.23 09D12 1.30E+07 3.50E-03 0.149 0.27 1.10E+07 7.40E-03 0.049 0.69 09H02 3.10E+06 1.30E-03 0.272 0.43 1.10E+07 2.70E-03 0.027 0.25 MH1 6.80E+06 3.50E-03 0.124 0.51 1.30E+07 5.60E-03 0.081 0.45 15E02 6.30E+06 3.90E-03 0.229 0.61 3.70E+06 9.50E-03 0.034 2.5 MH5 5.80E+06 4.10E-03 0.166 0.71 1.20E+07 6.00E-03 0.076 0.51 MH3 7.60E+06 6.50E-03 0.143 0.85 1.80E+07 1.10E-02 0.057 0.63 TTP 6.20E+07 7.30E-02 1.21 1.2 ND ND ND ND 11H02 6.10E+06 8.10E-03 1.04 1.3 1.70E+07 3.70E-02 0.332 2.1
TABLE-US-00007 TABLE 6 Amino acid sequences of VL-domains of unique, deimmunised anti-ERBB3 IgCs. Clone VL 15G11-DI1 DIQMTQSPSSLSASVGDRVTITCRASQEISSYLSWYQQKPGKAPKLLIYAASSLDTG VPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYDSTPYTFGGGTKVEIK (SEQ ID NO: 217) 15G11-DI2 DIQMTQSPSSLSASVGDRVTITCRASQEISTYLSWYQQKPGKAPKLLIYAASSLDTG VPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYDSTPYTFGGGTKVEIK (SEQ ID NO: 218) 15G11-DI3 DIQMTQSPSSLSASVGDRVTITCRASQEISSYLSWYQQKPGKAPKLLIYAASSLDTG VPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYDSSPYTFGGGTKVEIK (SEQ ID NO: 219) 15G11-DI4 DIQMTQSPSSLSASVGDRVTITCRASQEISTYLSWYQQKPGKAPKLLIYAASSLDTG VPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYDSSPYTFGGGTKVEIK (SEQ ID NO: 220) 15G11-DI5 DIQMTQSPSSLSASVGDRVTITCRASQEISSYLSWYQQKPGKAPKLLIYAASSLDTG VPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYDSTPYTFGGGTKVEIK (SEQ ID NO: 221) 15G11-DI6 DIQMTQSPSSLSASVGDRVTITCRASQEISTYLSWYQQKPGKAPKLLIYAASSLDTG VPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYDSTPYTFGGGTKVEIK (SEQ ID NO: 222) 15G11-DI7 DIQMTQSPSSLSASVGDRVTITCRASQEISSYLSWYQQKPGKAPKLLIYAASSLDTG VPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYDSSPYTFGGGTKVEIK (SEQ ID NO: 223) 15G11-DI8 DIQMTQSPSSLSASVGDRVTITCRASQEISTYLSWYQQKPGKAPKLLIYAASSLDTG VPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYDSSPYTFGGGTKVEIK (SEQ ID NO: 224) 15G11-DI9 DIQMTQSPSSLSASVGDRVTITCRASQEISTYLSWYQQKPGKAPKLLIYAASTLQSG VPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYDSSPLTFGGGTKVEIK (SEQ ID NO: 225) 15G11-DI10 DIQMTQSPSSLSASVGDRVTITCRASQEISSYLSWYQQKPGKAPKLLAYAASSLDSG VPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYDSTPYTFGGGTKVEIK (SEQ ID NO: 226) 15G11-DI11 DIQMTQSPSSLSASVGDRVTITCRASQEASSYLSWYQQKPGKAPKLLAYAASSLDSG VPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYDSSPYTFGGGTKVEIK (SEQ ID NO: 227)
TABLE-US-00008 TABLE 7 Amino acid sequences of VH-domains of unique, deimmunised anti-ERBB3 IgGs. Clone VH 15G11-DI1 QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYGMSWVRQAPGKGLEWVSTISDSGSYTYY PDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARELGDYDGFDYWGQGTLVTVSS (SEQ ID NO: 228) 15G11-DI2 QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYGMSWVRQAPGKGLEWVSTISDSGSYTYY PDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARELGDYDGFDYWGQGTLVTVSS (SEQ ID NO: 229) 15G11-DI3 QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYGMSWVRQAPGKGLEWVSTISDSGSYTYY PDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARELGDYDGFDYWGQGTLVTVSS (SEQ ID NO: 230) 15G11-DI4 QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYGMSWVRQAPGKGLEWVSTISDSGSYTYY PDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARELGDYDGFDYWGQGTLVTVSS (SEQ ID NO: 231) 15G11-DI5 QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYSMSWIRQAPGKGLEWVSTISDSGTYTYY PDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREWGDYDGFDFWGQGTLVTVSS (SEQ ID NO: 232) 15G11-DI6 QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYSMSWIRQAPGKGLEWVSTISDSGTYTYY PDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREWGDYDGFDFWGQGTLVTVSS (SEQ ID NO: 233) 15G11-DI7 QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYSMSWIRQAPGKGLEWVSTISDSGTYTYY PDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREWGDYDGFDFWGQGTLVTVSS (SEQ ID NO: 234) 15G11-DI8 QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYSMSWIRQAPGKGLEWVSTISDSGTYTYY PDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREWGDYDGFDFWGQGTLVTVSS (SEQ ID NO: 235) 15G11-DI9 QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYSMSWIRQAPGKGLEWVSTISDSGTYTYY PDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREWGDYDGFDFWGQGTLVTVSS (SEQ ID NO: 236) 15G11-DI10 QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYGMSWVRQAPGKGLEWVSTISDSGSYTYY PDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARELGDYDGFDYWGQGTLVTVSS (SEQ ID NO: 237) 15G11-DI11 QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYGMSWVRQAPGKGLEWVSTISDSGSYTYY PDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARELGDYDGFDYWGQGTLVTVSS (SEQ ID NO: 238)
TABLE-US-00009 TABLE 8 Examples of antibody variable region amino acid sequences. Antibody 15G11-DI9 heavy chain variable (VH) region QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYSMSWIRQAPGKGLEVVVSTISDSGTYTY YPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREWGDYDGFDFWGQGTLVTV SS (SEQ ID NO: 236) Antibody 15G11-DI9 light chain variable (VL) region DIQMTQSPSSLSASVGDRVTITCRASQEISTYLSVVYQQKPGKAPKLLIYAASTLQSGVPSR FSGSGSGTDFTLTISSLQPEDFATYYCLQYDSSPLTFGGGTKVEIK (SEQ ID NO: 225) Antibody 15G11-DI5 heavy chain variable (VH) region QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYSMSWIRQAPGKGLEVVVSTISDSGTYTY YPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREWGDYDGFDFWGQGTLVTV SS (SEQ ID NO: 232) Antibody 15G11-DI5 light chain variable (VL) region DIQMTQSPSSLSASVGDRVTITCRASQEISSYLSVVYQQKPGKAPKLLIYAASSLDTGVPSR FSGSGSGTDFTLTISSLQPEDFATYYCLQYDSTPYTFGGGTKVEIK (SEQ ID NO: 221) Antibody 15G11 heavy chain variable (VH) region QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYGMSWIRQAPGKGLEVVVSTISDSGSYTY YPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARELGDYDGFDYWGQGTLVTVS S (SEQ ID NO: 253) Antibody 15G11 light chain variable (VL) region DIQMTQSPSSLSASVGDRVTITCRASQEISSYLSVVYQQKPGKAPKLLIYAASSLDSGVPSR FSGSGSGTDFTLTISSLQPEDFATYYCLQYDSTPYTFGGGTKVEIK (SEQ ID NO: 254) Antibody 161309 heavy chain variable (VH) region QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYSMSWIRQAPGKGLEVVVSTISDSGTYTY YPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREWGDYDGFDFWGQGTLVTV SS (SEQ ID NO: 255) Antibody 161309 light chain variable (VL) region DIQMTQSPSSLSASVGDRVTITCRASQEISSYLSVVYQQKPGKAPKLLIYAASTLQSGVPSR FSGSGSGTDFTLTISSLQPEDFATYYCLQYDSTPLTFGGGTKVEIK (SEQ ID NO: 256)
TABLE-US-00010 TABLE 9 Examples of antibody Fc region amino acid sequences. Human IgG4 wild type ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSVVNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNVVYVDGVEVHNAKTKPREEQFNST YRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQ EGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 239) Human IgG4(S228P) ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSVVNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNVVYVDGVEVHNAKTKPREEQFNST YRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQ EGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 240) Human IgG1 wild type ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNVVYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEL TKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 241) Human IgG1-3M ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGA PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNVVYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEL TKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 242) Human IgG2 wild type ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSVVNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNVVYVDGVEVHNAKTKPREEQFNSTF RVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDISVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 243) Human IgG1 wild type "REEM" allotype ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNVVYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 244) Human IgG1-3M "REEM" allotype ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGA PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNVVYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 245)
TABLE-US-00011 TABLE 10 Examples of ERBB3 protein amino acid sequences. Human ERBB3 sequence MRANDALQVLGLLFSLARGSEVGNSQAVCPGTLNGLSVTGDAENQYQTLYKLYERCEVVM GNLEIVLTGHNADLSFLQWIREVTGYVLVAMNEFSTLPLPNLRVVRGTQVYDGKFAIFVM LNYNTNSSHALRQLRLTQLTEILSGGVYIEKNDKLCHMDTIDWRDIVRDRDAEIVVKDNG RSCPPCHEVCKGRCWGPGSEDCQTLTKTICAPQCNGHCFGPNPNQCCHDECAGGCSGPQD TDCFACRHFNDSGACVPRCPQPLVYNKLTFQLEPNPHTKYQYGGVCVASCPHNFVVDQTS CVRACPPDKMEVDKNGLKMCEPCGGLCPKACEGTGSGSRFQTVDSSNIDGFVNCTKILGN LDFLITGLNGDPWHKIPALDPEKLNVFRTVREITGYLNIQSWPPHMHNFSVESNLTTIGG RSLYNRGFSLLIMKNLNVTSLGFRSLKEISAGRIYISANRQLCYHHSLNWTKVLRGPTEE RLDIKHNRPRRDCVAEGKVCDPLCSSGGCWGPGPGQCLSCRNYSRGGVCVTHCNFLNGEP REFAHEAECFSCHPECQPMEGTATCNGSGSDTCAQCAHFRDGPHCVSSCPHGVLGAKGPI YKYPDVQNECRPCHENCTQGCKGPELQDCLGQTLVLIGKTHLTMALTVIAGLVVIFMMLG GTFLYWRGRRIQNKRAMRRYLERGESIEPLDPSEKANKVLARIFKETELRKLKVLGSGVF GTVHKGVWIPEGESIKIPVCIKVIEDKSGRQSFQAVTDHMLAIGSLDHAHIVRLLGLCPG SSLQLVTQYLPLGSLLDHVRQHRGALGPQLLLNWGVQIAKGMYYLEEHGMVHRNLAARNV LLKSPSQVQVADFGVADLLPPDDKQLLYSEAKTPIKWMALESIHFGKYTHQSDVWSYGVT VWELMTFGAEPYAGLRLAEVPDLLEKGERLAQPQICTIDVYMVMVKCWMIDENIRPTFKE LANEFTRMARDPPRYLVIKRESGPGIAPGPEPHGLTNKKLEEVELEPELDLDLDLEAEED NLATTTLGSALSLPVGTLNRPRGSQSLLSPSSGYMPMNQGNLGESCQESAVSGSSERCPR PVSLHPMPRGCLASESSEGHVTGSEAELQEKVSMCRSRSRSRSPRPRGDSAYHSQRHSLL TPVTPLSPPGLEEEDVNGYVMPDTHLKGTPSSREGTLSSVGLSSVLGTEEEDEDEEYEYM NRRRRHSPPHPPRPSSLEELGYEYMDVGSDLSASLGSTQSCPLHPVPIMPTAGTTPDEDY EYMNRQRDGGGPGGDYAAMGACPASEQGYEEMRAFQGPGHQAPHVHYARLKTLRSLEATD SAFDNPDYWHSRLFPKANAQRT (SEQ ID NO: 246) Rhesus monkey ERBB3 sequence MRANGALQVLGLLFNLARGSEVGNSQAVCPGTLNGLSVTGDAENQYQTLYKLYERCEVVM GNLEIVLTGHNADLSFLQWIREVTGYVLVAMNEFSTLPLPNLRVVRGTQVYDGKFAIFVM LNYNTNSSHALRQLRLTQLTEILSGGVYIEKNDKLCHMDTIDWKDIVRDQDAEIVVKDNG RSCPLCHEVCKGRCWGPGPEDCQTLTKTICAPQCNGHCFGPNPNQCCHDECAGGCSGPQD TDCFACRHFNDSGACVPRCPQPLVYNKLTFQLEPNPHTKYQYGGVCVASCPHNFVVDQTS CVRACPPDKMEVDKNGLKMCEPCGGLCPKACEGTGSGSRFQTVDSSNIDGFVNCTKILGN LDFLITGLNGDPWHKIPALDPEKLNVFRTVREITGYLNIQSWPPHMYNFSVESNLTTIGG RSLYNRGFSLLIMKNLNVTSLGFRSLKEISAGRIYISANRQLCYHHSLNWTKVLRGPTEE RLDIKHNRPRRDCVAEGKVCDPLCSSGGCWGPGPGQCLSCRNYSRGGVCVTHCNFLNGEP REFAHEAECFSCHPECQPMEGTATCNGSGSDTCAQCAHFRDGPHCVSSCPHGVLGAKGPI YKYPDVQNECRPCHENCTQGCKGPELQDCLGQTLVLIGKTHLTMALTVIAGLVVIFMMLG GTFLYWRGRRIQNKRAMRRYLERGESIEPLDPSEKANKVLARIFKETELRKLKVLGSGVF GTVHKGVWIPEGESIKIPVCIKIIEDKSGRQSFQAVTDHMLAIGSLDHAHIVRLLGLCPG SSLQLVTQYLPLGSLLDHVRQHRGALGPQLLLNWGVQIAKGMYYLEEHGMVHRNLAARNV LLKSPSQVQVADFGVADLLPPDDKQLLYSEAKTPIKWMALESIHFGKYTHQSDVWSYGVT VWELMTFGAEPYAGLRLAEVPDLLEKGERLAQPQICTIDVYMVMVKCWMIDENIRPTFKE LANEFTRMARDPPRYLVIKRESGPGIAPGPEPHGLTNKKLEEVELEPELDLDLDLEAEED NLATTTLGSALSLPVGTLNRPRGSQSLLSPSSGYMPMNQGNLGEACQESAVSGSSEWCPR PVSLHPMPRGCLASESSEGHVTGSEAELQEKVSTCRSRSRSRSPRPRGDSAYHSQRHSLL TPVTPLSPPGLEEEDVNGYVMPDTHLKGTPSSREGTLSSVGLSSVLGTEEEDEDEEYEYM NRRRRHSPPRPPRPSSLEELGYEYMDVGSDLSASLGSTQSCPLHPVPVMPTAGTTPDEDY EYMNRQRGGSGPGGDYAAMGACPASEQGYEEMRAFQGPGHQAPHVHYAHLKTLRSLEATD SAFDNPDYWHSRLFPKANAQRT (SEQ ID NO: 247)
Sequence CWU
1
1
263110PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR1misc_feature(8)..(8)Xaa is Gly or any
other amino acid 1Gly Phe Thr Phe Ser Asp Tyr Xaa Met Ser1
5 10219PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2misc_feature(7)..(7)Xaa is
Gly or any other amino acidmisc_feature(9)..(9)Xaa is Thr or a
conservative substitution of Thrmisc_feature(10)..(10)Xaa is Tyr or
any other amino acidmisc_feature(11)..(11)Xaa is Thr or any other amino
acidmisc_feature(14)..(14)Xaa is Pro or any other amino
acidmisc_feature(16)..(16)Xaa is Asn or a conservative substitution of
Asn 2Val Ser Thr Ile Ser Asp Xaa Gly Xaa Xaa Xaa Tyr Tyr Xaa Asp Xaa1
5 10 15Val Lys
Gly310PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3misc_feature(1)..(1)Xaa is Glu or any
other amino acidmisc_feature(2)..(2)Xaa is Trp or any other amino
acidmisc_feature(5)..(5)Xaa is Tyr or any other amino
acidmisc_feature(8)..(8)Xaa is Phe or any other amino
acidmisc_feature(10)..(10)Xaa is Tyr or any other amino acid 3Xaa Xaa Gly
Asp Xaa Asp Gly Xaa Asp Xaa1 5
10410PRTArtificial Sequence24C05 murine/humanized antibody HCDR1 4Gly Phe
Thr Phe Ser Asp Tyr Ala Met Ser1 5
10519PRTArtificial Sequence24C05 murine/humanized antibody HCDR2 5Val Ser
Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Asn1 5
10 15Val Lys Gly610PRTArtificial
Sequence24C05 murine/humanized antibody HCDR3 6Glu Trp Gly Asp Tyr Asp
Gly Phe Asp Tyr1 5 10711PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
LCDR1misc_feature(5)..(5)Xaa is Glu or any other amino
acidmisc_feature(8)..(8)Xaa is Gly or a conservative substitution of
Glymisc_feature(11)..(11)Xaa is Ser or a conservative substitution of
Ser 7Arg Ala Ser Gln Xaa Ile Ser Xaa Tyr Leu Xaa1 5
1087PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR2misc_feature(1)..(1)Xaa is Ala or any
other amino acidmisc_feature(4)..(4)Xaa is Thr or a conservative
substitution of Thrmisc_feature(6)..(6)Xaa is Asp any other amino
acid 8Xaa Ala Ser Xaa Leu Xaa Ser1 599PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
LCDR3misc_feature(1)..(1)Xaa is Leu or any other amino
acidmisc_feature(3)..(3)Xaa is Tyr or any other amino
acidmisc_feature(4)..(4)Xaa is Asp or any other amino
acidmisc_feature(6)..(6)Xaa is Tyr or any other amino
acidmisc_feature(7)..(7)Xaa is Pro or any other amino
acidmisc_feature(8)..(8)Xaa is Tyr or any other amino acid 9Xaa Gln Xaa
Xaa Ser Xaa Xaa Xaa Thr1 51011PRTArtificial Sequence24C05
murine/humanized antibody LCDR1 10Arg Ala Ser Gln Glu Ile Ser Gly Tyr Leu
Ser1 5 10117PRTArtificial Sequence24C05
murine/humanized antibody LCDR2 11Ala Ala Ser Thr Leu Asp Ser1
5129PRTArtificial Sequence24C05 murine/humanized antibody LCDR3 12Leu
Gln Tyr Asp Ser Tyr Pro Tyr Thr1 51310PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR1 13Gly Phe Thr Phe Ser Asp Tyr Gly Met Ser1 5
101419PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 14Val Ser Thr Ile Ser Asp Ser Gly Ser
Tyr Ile Tyr Tyr Ala Asp Ser1 5 10
15Val Lys Gly1510PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR3 15Glu Trp Gly Asp Tyr Asp
Gly Phe Asp Phe1 5 101611PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
LCDR1 16Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu Ser1 5
10177PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR2 17Ala Ala Ser Ser Leu Gln Ser1
5189PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR3 18Leu Gln Tyr Asp Ser Thr Pro Leu Thr1
51919PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 19Val Ser Thr Ile Ser Asp Ser Gly Ser
Tyr Thr Tyr Tyr Pro Asp Ser1 5 10
15Val Lys Gly2010PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR3 20Glu Leu Gly Asp Tyr Asp
Gly Phe Asp Tyr1 5 102111PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
LCDR1 21Arg Ala Ser Gln Glu Ile Ser Ser Tyr Leu Ser1 5
10227PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR2 22Ala Ala Ser Ser Leu Asp Ser1
5239PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR3 23Leu Gln Tyr Asp Ser Thr Pro Tyr Thr1
52410PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR1 24Gly Phe Thr Phe Ser Asp Tyr Ser Met
Ser1 5 102519PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 25Val Ser Thr Ile Ser Asp Ser Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser1
5 10 15Val Lys
Gly267PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR2 26Ala Ala Ser Thr Leu Gln Ser1
52710PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 27Glu Trp Gly Asp Tyr Asp Gly Phe Asp
His1 5 102819PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 28Val Ser Thr Ile Ser Asp Gly Gly Ser Tyr Thr Tyr Tyr Ala Asp Ser1
5 10 15Val Lys
Gly2910PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 29Glu Trp Gly Asp Tyr Asp Gly Phe Asp
Glu1 5 103011PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
LCDR1 30Arg Ala Ser Gln Ser Ile Ser Gly Tyr Leu Ser1 5
103119PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 31Val Ser Thr Ile Ser Asp Gly Gly Ser
Tyr Thr Tyr Tyr Ala Asp Asn1 5 10
15Val Lys Gly3211PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion consensus LCDR1 32Arg Ala Ser
Gln Glu Ile Ser Gly Tyr Leu Ser1 5
10337PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion consensus LCDR2 33Ala Ala Ser Thr Leu Asp
Ser1 5349PRTArtificial Sequenceanti-ERBB3 antibody molecule
or antigen-binding portion consensus LCDR3 34Leu Gln Tyr Asp Ser Tyr
Pro Tyr Thr1 53510PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion consensus HCDR1 35Gly Phe Thr
Phe Ser Asp Tyr Ala Met Ser1 5
103619PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion consensus HCDR2 36Val Ser Thr Ile Ser Asp
Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Asn1 5
10 15Val Lys Gly3710PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion consensus HCDR3 37Glu
Trp Gly Asp Tyr Asp Gly Phe Asp Tyr1 5
103810PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR1misc_feature(8)..(8)Xaa is Glu, Gly,
His, Asn, Arg, Ser, Thr, Gln or Val 38Gly Phe Thr Phe Ser Asp Tyr
Xaa Met Ser1 5 103919PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2misc_feature(7)..(7)Xaa is Gly, Ser or Aspmisc_feature(9)..(9)Xaa is
Thr or Sermisc_feature(10)..(10)Xaa is Tyr or
Thrmisc_feature(11)..(11)Xaa is Thr or Ilemisc_feature(14)..(14)Xaa is
Pro or Alamisc_feature(16)..(16)Xaa is Asn or Ser 39Val Ser Thr Ile Ser
Asp Xaa Gly Xaa Xaa Xaa Tyr Tyr Xaa Asp Xaa1 5
10 15Val Lys Gly4010PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3misc_feature(1)..(1)Xaa is Glu or Metmisc_feature(2)..(2)Xaa is Trp,
Phe, Leu, Met, Gln or Tyrmisc_feature(5)..(5)Xaa is Tyr, Ala, Asp, Glu,
His, Leu, Met, Asn, Gln, Ser, Thr or Trpmisc_feature(8)..(8)Xaa is
Phe, Ile, Leu, Trp or Tyrmisc_feature(10)..(10)Xaa is Tyr, Ala, Asp, Glu,
Phe, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Val or Trp 40Xaa
Xaa Gly Asp Xaa Asp Gly Xaa Asp Xaa1 5
104110PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR1misc_feature(8)..(8)Xaa is Gly or Ser
41Gly Phe Thr Phe Ser Asp Tyr Xaa Met Ser1 5
104219PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2misc_feature(7)..(7)Xaa is Gly or
Sermisc_feature(10)..(10)Xaa is Tyr or Thrmisc_feature(11)..(11)Xaa is
Thr or Ilemisc_feature(14)..(14)Xaa is Pro or Ala 42Val Ser Thr Ile Ser
Asp Xaa Gly Ser Xaa Xaa Tyr Tyr Xaa Asp Ser1 5
10 15Val Lys Gly4310PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3misc_feature(2)..(2)Xaa is Trp, Leu or Tyrmisc_feature(10)..(10)Xaa
is Tyr, Glu, Phe, His or Asn 43Glu Xaa Gly Asp Tyr Asp Gly Phe Asp Xaa1
5 104411PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion
LCDR1misc_feature(5)..(5)Xaa is Glu, Ser, Ile or
Asnmisc_feature(8)..(8)Xaa is Gly or Sermisc_feature(11)..(11)Xaa is Ser
or Asn 44Arg Ala Ser Gln Xaa Ile Ser Xaa Tyr Leu Xaa1 5
10457PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR2misc_feature(1)..(1)Xaa is Ala or
Glumisc_feature(4)..(4)Xaa is Thr, Ser or Asnmisc_feature(6)..(6)Xaa is
Asp, His, Lys or Gln 45Xaa Ala Ser Xaa Leu Xaa Ser1
5469PRTArtificial Sequenceanti-ERBB3 antibody molecule or antigen-binding
portion LCDR3misc_feature(1)..(1)Xaa is Leu or
Glnmisc_feature(3)..(3)Xaa is Tyr or Sermisc_feature(4)..(4)Xaa is Asp or
Tyrmisc_feature(6)..(6)Xaa is Tyr or Thrmisc_feature(7)..(7)Xaa is Pro or
Hismisc_feature(8)..(8)Xaa is Tyr or Leu 46Xaa Gln Xaa Xaa Ser Xaa Xaa
Xaa Thr1 54711PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion LCDR1misc_feature(5)..(5)Xaa is
Glu or Sermisc_feature(8)..(8)Xaa is Gly or Sermisc_feature(11)..(11)Xaa
is Ser or Asn 47Arg Ala Ser Gln Xaa Ile Ser Xaa Tyr Leu Xaa1
5 10487PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion LCDR2misc_feature(4)..(4)Xaa is
Thr or Sermisc_feature(6)..(6)Xaa is Asp or Gln 48Ala Ala Ser Xaa Leu Xaa
Ser1 5499PRTArtificial Sequenceanti-ERBB3 antibody molecule
or antigen-binding portion LCDR3misc_feature(4)..(4)Xaa is Asp or
Tyrmisc_feature(8)..(8)Xaa is Tyr or Leu 49Leu Gln Tyr Xaa Ser Thr Pro
Xaa Thr1 55011PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion LCDR1 50Arg Ala Ser Gln Ser Ile
Ser Ser Tyr Leu Asn1 5
105110PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 51Glu Tyr Gly Asp Tyr Asp Gly Phe Asp
Tyr1 5 105211PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
LCDR1 52Arg Ala Ser Gln Ile Ile Ser Ser Tyr Leu Ser1 5
10539PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR3 53Leu Gln Tyr Tyr Ser Thr Pro Leu Thr1
55419PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 54Val Ser Thr Ile Ser Asp Ser Gly Ser
Tyr Thr Tyr Tyr Ala Asp Ser1 5 10
15Val Lys Gly5510PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR3 55Glu Trp Gly Asp Tyr Asp
Gly Phe Asp Asn1 5 105619PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 56Val Ser Thr Ile Ser Asp Ser Gly Thr Thr Ile Tyr Tyr Ala Asp Asn1
5 10 15Val Lys
Gly5719PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 57Val Ser Thr Ile Ser Asp Gly Gly Ser
Tyr Thr Tyr Tyr Pro Asp Ser1 5 10
15Val Lys Gly5819PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 58Val Ser Thr Ile Ser Asp
Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser1 5
10 15Val Lys Gly5910PRTMus musculus 59Gly Phe Thr Phe
Ser Asp Tyr Ala Met Ser1 5 10605PRTMus
musculus 60Asp Tyr Ala Met Ser1 5617PRTMus musculus 61Gly
Phe Thr Phe Ser Asp Tyr1 5628PRTMus musculus 62Gly Phe Thr
Phe Ser Asp Tyr Ala1 56312PRTMus musculus 63Ala Ser Gly Phe
Thr Phe Ser Asp Tyr Ala Met Ser1 5
10646PRTMus musculus 64Ser Asp Tyr Ala Met Ser1 56519PRTMus
musculus 65Val Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp
Asn1 5 10 15Val Lys
Gly6617PRTMus musculus 66Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro
Asp Asn Val Lys1 5 10
15Gly676PRTMus musculus 67Ser Asp Gly Gly Thr Tyr1
5688PRTMus musculus 68Ile Ser Asp Gly Gly Thr Tyr Thr1
56916PRTMus musculus 69Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp
Asn Val Lys Gly1 5 10
157010PRTMus musculus 70Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr1
5 107112PRTMus musculus 71Val Ser Thr Ile Ser Asp
Gly Gly Thr Tyr Thr Tyr1 5 107210PRTMus
musculus 72Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr1 5
107312PRTMus musculus 73Ala Arg Glu Trp Gly Asp Tyr Asp Gly
Phe Asp Tyr1 5 10749PRTMus musculus 74Glu
Trp Gly Asp Tyr Asp Gly Phe Asp1 57511PRTMus musculus 75Ala
Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp1 5
107611PRTMus musculus 76Arg Ala Ser Gln Glu Ile Ser Gly Tyr Leu Ser1
5 10776PRTMus musculus 77Gln Glu Ile Ser Gly
Tyr1 5788PRTMus musculus 78Ala Ser Gln Glu Ile Ser Gly Tyr1
5798PRTMus musculus 79Ile Ser Gly Tyr Leu Ser Trp Tyr1
5807PRTMus musculus 80Ala Ala Ser Thr Leu Asp Ser1
58110PRTMus musculus 81Leu Leu Ile Tyr Ala Ala Ser Thr Leu Asp1
5 10829PRTMus musculus 82Leu Gln Tyr Asp Ser Tyr
Pro Tyr Thr1 5836PRTMus musculus 83Tyr Asp Ser Tyr Pro Tyr1
5848PRTMus musculus 84Leu Gln Tyr Asp Ser Tyr Pro Tyr1
585119PRTArtificial Sequenceh24C05-VH IGHV3-11 85Gln Val Gln Leu
Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5
10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
Phe Thr Phe Ser Asp Tyr 20 25
30Ala Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45Ser Thr Ile Ser Asp Gly Gly Thr
Tyr Thr Tyr Tyr Pro Asp Asn Val 50 55
60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65
70 75 80Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp
Tyr Trp Gly Gln Gly 100 105
110Thr Leu Val Thr Val Ser Ser 11586119PRTArtificial SequenceVH
graft IGHV3-11 86Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro
Gly Gly1 5 10 15Ser Leu
Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20
25 30Ala Met Ser Trp Ile Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40
45Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Asn Val 50
55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ala Lys Asn Ser Leu Tyr65 70 75
80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95Ala Arg
Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100
105 110Thr Leu Val Thr Val Ser Ser
11587107PRTArtificial Sequenceh24C05-VL IGKV1-16 87Asp Ile Gln Met Thr
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5
10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln
Glu Ile Ser Gly Tyr 20 25
30Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile
35 40 45Tyr Ala Ala Ser Thr Leu Asp Ser
Gly Val Pro Ser Arg Phe Ser Gly 50 55
60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80Glu Asp Phe Ala Thr
Tyr Tyr Cys Leu Gln Tyr Asp Ser Tyr Pro Tyr 85
90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 10588107PRTArtificial SequenceVL graft
IGKV1-39 88Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val
Gly1 5 10 15Asp Arg Val
Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Gly Tyr 20
25 30Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys
Ala Pro Lys Leu Leu Ile 35 40
45Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly 50
55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Ser Leu Gln Pro65 70 75
80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Tyr
Pro Tyr 85 90 95Thr Phe
Gly Gly Gly Thr Lys Val Glu Ile Lys 100
1058911PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR1 89Arg Ala Ser Gln Ser Ile Ser Gly Tyr
Leu Asn1 5 109011PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
LCDR1 90Arg Ala Ser Gln Glu Ile Ser Ser Tyr Leu Asn1 5
109111PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR1 91Arg Ala Ser Gln Glu Ile Ser Gly Tyr
Leu Asn1 5 109211PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
LCDR1 92Arg Ala Ser Gln Asn Ile Ser Ser Tyr Leu Ser1 5
10937PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR2 93Ala Ala Ser Asn Leu Gln Ser1
5947PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR2 94Ala Ala Ser Ser Leu His Ser1
5957PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR2 95Glu Ala Ser Ser Leu Asp Ser1
5967PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR2 96Ala Ala Ser Ser Leu Lys Ser1
5977PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR2 97Glu Ala Ser Ser Leu Gln Ser1
5989PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR3 98Leu Gln Tyr Tyr Ser Tyr Pro Tyr Thr1
5999PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR3 99Gln Gln Tyr Asp Ser Tyr Pro Tyr Thr1
51009PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR3 100Leu Gln Tyr Asp Ser Tyr Pro Leu
Thr1 51019PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion LCDR3 101Gln Gln Tyr Asp Ser Thr
Pro Tyr Thr1 51029PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion LCDR3 102Leu Gln Ser Asp Ser Thr
Pro Tyr Thr1 51039PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion LCDR3 103Leu Gln Ser Asp Ser Thr
Pro Leu Thr1 51049PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion LCDR3 104Gln Gln Tyr Asp Ser Tyr
Pro Leu Thr1 51059PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion LCDR3 105Leu Gln Tyr Tyr Ser Tyr
Pro Leu Thr1 51069PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion LCDR3 106Leu Gln Tyr Tyr Ser Thr
Pro Tyr Thr1 51079PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion LCDR3 107Gln Gln Tyr Asp Ser Thr
Pro Leu Thr1 51089PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion LCDR3 108Leu Gln Tyr Asp Ser Tyr
His Leu Thr1 510910PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR1 109Gly Phe Thr
Phe Ser Asp Tyr Ala Met Ser1 5
1011010PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR1 110Gly Phe Thr Phe Ser Asp Tyr Glu Met
Ser1 5 1011110PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR1 111Gly Phe Thr Phe Ser Asp Tyr His Met Ser1 5
1011210PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR1 112Gly Phe Thr Phe Ser Asp Tyr Asn Met
Ser1 5 1011310PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR1 113Gly Phe Thr Phe Ser Asp Tyr Gln Met Ser1 5
1011410PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR1 114Gly Phe Thr Phe Ser Asp Tyr Arg Met
Ser1 5 1011510PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR1 115Gly Phe Thr Phe Ser Asp Tyr Thr Met Ser1 5
1011610PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR1 116Gly Phe Thr Phe Ser Asp Tyr Val Met
Ser1 5 1011719PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 117Val Gly Thr Ile Ser Asp Gly Gly Thr Thr Ile Tyr Tyr Ala Asp Asn1
5 10 15Val Lys
Gly11819PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 118Val Ser Thr Ile Ser Asp Asp Gly Ser
Thr Thr Tyr Tyr Ala Asp Ser1 5 10
15Val Lys Gly11919PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 119Val Ser Thr Ile Ser Asp
Gly Gly Ser Thr Ile Tyr Tyr Ala Asp Asn1 5
10 15Val Lys Gly12019PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 120Val Ser Thr
Ile Ser Asp Gly Gly Ser Thr Ile Tyr Tyr Ala Asp Ser1 5
10 15Val Lys Gly12119PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 121Val Ser Thr Ile Ser Asp Gly Gly Ser Thr Ile Tyr Tyr Pro Asp Asn1
5 10 15Val Lys
Gly12219PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 122Val Ser Thr Ile Ser Asp Gly Gly Ser
Thr Ile Tyr Tyr Pro Asp Ser1 5 10
15Val Lys Gly12319PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 123Val Ser Thr Ile Ser Asp
Gly Gly Ser Thr Thr Tyr Tyr Ala Asp Asn1 5
10 15Val Lys Gly12419PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 124Val Ser Thr
Ile Ser Asp Gly Gly Ser Thr Thr Tyr Tyr Ala Asp Ser1 5
10 15Val Lys Gly12519PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 125Val Ser Thr Ile Ser Asp Gly Gly Ser Thr Thr Tyr Tyr Pro Asp Asn1
5 10 15Val Lys
Gly12619PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 126Val Ser Thr Ile Ser Asp Gly Gly Ser
Thr Thr Tyr Tyr Pro Asp Ser1 5 10
15Val Lys Gly12719PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 127Val Ser Thr Ile Ser Asp
Gly Gly Ser Tyr Ile Tyr Tyr Ala Asp Asn1 5
10 15Val Lys Gly12819PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 128Val Ser Thr
Ile Ser Asp Gly Gly Ser Tyr Ile Tyr Tyr Ala Asp Ser1 5
10 15Val Lys Gly12919PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 129Val Ser Thr Ile Ser Asp Gly Gly Ser Tyr Ile Tyr Tyr Pro Asp Asn1
5 10 15Val Lys
Gly13019PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 130Val Ser Thr Ile Ser Asp Gly Gly Ser
Tyr Ile Tyr Tyr Pro Asp Ser1 5 10
15Val Lys Gly13119PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 131Val Ser Thr Ile Ser Asp
Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Asn1 5
10 15Val Lys Gly13219PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 132Val Ser Thr
Ile Ser Asp Gly Gly Thr Thr Ile Tyr Tyr Ala Asp Asn1 5
10 15Val Lys Gly13319PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 133Val Ser Thr Ile Ser Asp Gly Gly Thr Thr Ile Tyr Tyr Ala Asp Ser1
5 10 15Val Lys
Gly13419PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 134Val Ser Thr Ile Ser Asp Gly Gly Thr
Thr Ile Tyr Tyr Pro Asp Ser1 5 10
15Val Lys Gly13519PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 135Val Ser Thr Ile Ser Asp
Gly Gly Thr Thr Thr Tyr Tyr Ala Asp Asn1 5
10 15Val Lys Gly13619PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 136Val Ser Thr
Ile Ser Asp Gly Gly Thr Thr Thr Tyr Tyr Ala Asp Ser1 5
10 15Val Lys Gly13719PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 137Val Ser Thr Ile Ser Asp Gly Gly Thr Thr Thr Tyr Tyr Pro Asp Asn1
5 10 15Val Lys
Gly13819PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 138Val Ser Thr Ile Ser Asp Gly Gly Thr
Thr Thr Tyr Tyr Pro Asp Ser1 5 10
15Val Lys Gly13919PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 139Val Ser Thr Ile Ser Asp
Gly Gly Thr Tyr Ile Tyr Tyr Ala Asp Asn1 5
10 15Val Lys Gly14019PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 140Val Ser Thr
Ile Ser Asp Gly Gly Thr Tyr Ile Tyr Tyr Ala Asp Ser1 5
10 15Val Lys Gly14119PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 141Val Ser Thr Ile Ser Asp Gly Gly Thr Tyr Ile Tyr Tyr Pro Asp Asn1
5 10 15Val Lys
Gly14219PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 142Val Ser Thr Ile Ser Asp Gly Gly Thr
Tyr Ile Tyr Tyr Pro Asp Ser1 5 10
15Val Lys Gly14319PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 143Val Ser Thr Ile Ser Asp
Gly Gly Thr Tyr Thr Tyr Tyr Ala Asp Asn1 5
10 15Val Lys Gly14419PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 144Val Ser Thr
Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Ala Asp Ser1 5
10 15Val Lys Gly14519PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 145Val Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Asn1
5 10 15Val Lys
Gly14619PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 146Val Ser Thr Ile Ser Asp Gly Gly Thr
Tyr Thr Tyr Tyr Pro Asp Ser1 5 10
15Val Lys Gly14719PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 147Val Ser Thr Ile Ser Asp
Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Asn1 5
10 15Val Lys Gly14819PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 148Val Ser Thr
Ile Ser Asp Ser Gly Ser Thr Ile Tyr Tyr Pro Asp Asn1 5
10 15Val Lys Gly14919PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 149Val Ser Thr Ile Ser Asp Ser Gly Ser Thr Ile Tyr Tyr Pro Asp Ser1
5 10 15Val Lys
Gly15019PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 150Val Ser Thr Ile Ser Asp Ser Gly Ser
Thr Thr Tyr Tyr Ala Asp Asn1 5 10
15Val Lys Gly15119PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 151Val Ser Thr Ile Ser Asp
Ser Gly Ser Thr Thr Tyr Tyr Ala Asp Ser1 5
10 15Val Lys Gly15219PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 152Val Ser Thr
Ile Ser Asp Ser Gly Ser Thr Thr Tyr Tyr Pro Asp Asn1 5
10 15Val Lys Gly15319PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 153Val Ser Thr Ile Ser Asp Ser Gly Ser Thr Thr Tyr Tyr Pro Asp Ser1
5 10 15Val Lys
Gly15419PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 154Val Ser Thr Ile Ser Asp Ser Gly Ser
Tyr Ile Tyr Tyr Ala Asp Asn1 5 10
15Val Lys Gly15519PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 155Val Ser Thr Ile Ser Asp
Ser Gly Ser Tyr Ile Tyr Tyr Pro Asp Asn1 5
10 15Val Lys Gly15619PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 156Val Ser Thr
Ile Ser Asp Ser Gly Ser Tyr Ile Tyr Tyr Pro Asp Ser1 5
10 15Val Lys Gly15719PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 157Val Ser Thr Ile Ser Asp Ser Gly Ser Tyr Thr Tyr Tyr Ala Asp Asn1
5 10 15Val Lys
Gly15819PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 158Val Ser Thr Ile Ser Asp Ser Gly Ser
Tyr Thr Tyr Tyr Pro Asp Asn1 5 10
15Val Lys Gly15919PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 159Val Ser Thr Ile Ser Asp
Ser Gly Thr Thr Ile Tyr Tyr Ala Asp Ser1 5
10 15Val Lys Gly16019PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 160Val Ser Thr
Ile Ser Asp Ser Gly Thr Thr Thr Tyr Tyr Ala Asp Asn1 5
10 15Val Lys Gly16119PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 161Val Ser Thr Ile Ser Asp Ser Gly Thr Thr Thr Tyr Tyr Ala Asp Ser1
5 10 15Val Lys
Gly16219PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 162Val Ser Thr Ile Ser Asp Ser Gly Thr
Thr Thr Tyr Tyr Pro Asp Asn1 5 10
15Val Lys Gly16319PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 163Val Ser Thr Ile Ser Asp
Ser Gly Thr Thr Thr Tyr Tyr Pro Asp Ser1 5
10 15Val Lys Gly16419PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 164Val Ser Thr
Ile Ser Asp Ser Gly Thr Tyr Ile Tyr Tyr Ala Asp Asn1 5
10 15Val Lys Gly16519PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 165Val Ser Thr Ile Ser Asp Ser Gly Thr Tyr Ile Tyr Tyr Ala Asp Ser1
5 10 15Val Lys
Gly16619PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 166Val Ser Thr Ile Ser Asp Ser Gly Thr
Tyr Ile Tyr Tyr Pro Asp Asn1 5 10
15Val Lys Gly16719PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 167Val Ser Thr Ile Ser Asp
Ser Gly Thr Tyr Ile Tyr Tyr Pro Asp Ser1 5
10 15Val Lys Gly16819PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 168Val Ser Thr
Ile Ser Asp Ser Gly Thr Tyr Thr Tyr Tyr Ala Asp Asn1 5
10 15Val Lys Gly16919PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 169Val Ser Thr Ile Ser Asp Ser Gly Thr Tyr Thr Tyr Tyr Ala Asp Ser1
5 10 15Val Lys
Gly17019PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 170Val Ser Thr Ile Ser Asp Ser Gly Thr
Tyr Thr Tyr Tyr Pro Asp Asn1 5 10
15Val Lys Gly17119PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 171Val Ser Thr Ile Ser Asn
Ser Gly Thr Tyr Thr Tyr Tyr Ala Asp Ser1 5
10 15Val Lys Gly17219PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 172Val Ser Thr
Ile Ser Ser Gly Gly Ser Tyr Ile Tyr Tyr Pro Asp Ser1 5
10 15Val Lys Gly17319PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 173Val Ser Thr Ile Ser Ser Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser1
5 10 15Val Lys
Gly17419PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 174Val Ser Thr Ile Ser Ser Ser Gly Ser
Tyr Thr Tyr Tyr Ala Asp Asn1 5 10
15Val Lys Gly17519PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 175Val Ser Thr Ile Ser Ser
Ser Gly Thr Thr Thr Tyr Tyr Ala Asp Ser1 5
10 15Val Lys Gly17619PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 176Val Ser Thr
Ile Ser Ser Ser Gly Thr Tyr Ile Tyr Tyr Ala Asp Asn1 5
10 15Val Lys Gly17719PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR2 177Val Ser Tyr Ile Ser Asp Gly Gly Ser Tyr Ile Tyr Tyr Ala Asp Ser1
5 10 15Val Lys
Gly17819PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR2 178Val Ser Tyr Ile Ser Asp Gly Gly Ser
Tyr Thr Tyr Tyr Ala Asp Asn1 5 10
15Val Lys Gly17919PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion HCDR2 179Val Ser Tyr Ile Ser Asp
Gly Gly Thr Thr Ile Tyr Tyr Ala Asp Ser1 5
10 15Val Lys Gly18019PRTArtificial Sequenceanti-ERBB3
antibody molecule or antigen-binding portion HCDR2 180Val Ser Tyr
Ile Ser Asp Ser Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser1 5
10 15Val Lys Gly18110PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 181Glu Phe Gly Asp Tyr Asp Gly Phe Asp Phe1 5
1018210PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 182Glu Phe Gly Asp Tyr Asp Gly Phe Asp
Tyr1 5 1018310PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 183Glu Leu Gly Asp Tyr Asp Gly Trp Asp Tyr1 5
1018410PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 184Glu Met Gly Asp Tyr Asp Gly Phe Asp
Tyr1 5 1018510PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 185Glu Gln Gly Asp Tyr Asp Gly Phe Asp Ile1 5
1018610PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 186Glu Trp Gly Asp Ala Asp Gly Phe Asp
Tyr1 5 1018710PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 187Glu Trp Gly Asp Asp Asp Gly Phe Asp Tyr1 5
1018810PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 188Glu Trp Gly Asp Glu Asp Gly Phe Asp
Tyr1 5 1018910PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 189Glu Trp Gly Asp His Asp Gly Phe Asp Tyr1 5
1019010PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 190Glu Trp Gly Asp Leu Asp Gly Phe Asp
Tyr1 5 1019110PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 191Glu Trp Gly Asp Met Asp Gly Phe Asp Arg1 5
1019210PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 192Glu Trp Gly Asp Met Asp Gly Phe Asp
Tyr1 5 1019310PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 193Glu Trp Gly Asp Asn Asp Gly Phe Asp Tyr1 5
1019410PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 194Glu Trp Gly Asp Gln Asp Gly Phe Asp
Tyr1 5 1019510PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 195Glu Trp Gly Asp Ser Asp Gly Phe Asp Tyr1 5
1019610PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 196Glu Trp Gly Asp Thr Asp Gly Phe Asp
Tyr1 5 1019710PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 197Glu Trp Gly Asp Trp Asp Gly Phe Asp Tyr1 5
1019810PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 198Glu Trp Gly Asp Tyr Asp Gly Cys Asp
Tyr1 5 1019910PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 199Glu Trp Gly Asp Tyr Asp Gly Phe Asp Ala1 5
1020010PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 200Glu Trp Gly Asp Tyr Asp Gly Phe Asp
Asp1 5 1020110PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 201Glu Trp Gly Asp Tyr Asp Gly Phe Asp Ile1 5
1020210PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 202Glu Trp Gly Asp Tyr Asp Gly Phe Asp
Lys1 5 1020310PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 203Glu Trp Gly Asp Tyr Asp Gly Phe Asp Leu1 5
1020410PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 204Glu Trp Gly Asp Tyr Asp Gly Phe Asp
Met1 5 1020510PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 205Glu Trp Gly Asp Tyr Asp Gly Phe Asp Gln1 5
1020610PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 206Glu Trp Gly Asp Tyr Asp Gly Phe Asp
Arg1 5 1020710PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 207Glu Trp Gly Asp Tyr Asp Gly Phe Asp Ser1 5
1020810PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 208Glu Trp Gly Asp Tyr Asp Gly Phe Asp
Val1 5 1020910PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 209Glu Trp Gly Asp Tyr Asp Gly Phe Asp Trp1 5
1021010PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 210Glu Trp Gly Asp Tyr Asp Gly Phe Asp
Tyr1 5 1021110PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 211Glu Trp Gly Asp Tyr Asp Gly Phe His Tyr1 5
1021210PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 212Glu Trp Gly Asp Tyr Asp Gly Ile Asp
Tyr1 5 1021310PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 213Glu Trp Gly Asp Tyr Asp Gly Leu Asp Tyr1 5
1021410PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 214Glu Trp Gly Asp Tyr Asp Gly Trp Asp
Tyr1 5 1021510PRTArtificial
Sequenceanti-ERBB3 antibody molecule or antigen-binding portion
HCDR3 215Glu Trp Gly Asp Tyr Asp Gly Tyr Asp Tyr1 5
1021610PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion HCDR3 216Met Trp Gly Asp Tyr Asp Gly Phe Asp
Tyr1 5 10217107PRTArtificial
Sequence15G11-DI1 VL 217Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser
Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Ser Tyr
20 25 30Leu Ser Trp Tyr Gln Gln Lys
Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40
45Tyr Ala Ala Ser Ser Leu Asp Thr Gly Val Pro Ser Arg Phe Ser
Gly 50 55 60Ser Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70
75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr
Asp Ser Thr Pro Tyr 85 90
95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100
105218107PRTArtificial Sequence15G11-DI2 VL 218Asp Ile Gln Met Thr
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5
10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln
Glu Ile Ser Thr Tyr 20 25
30Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45Tyr Ala Ala Ser Ser Leu Asp Thr
Gly Val Pro Ser Arg Phe Ser Gly 50 55
60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80Glu Asp Phe Ala Thr
Tyr Tyr Cys Leu Gln Tyr Asp Ser Thr Pro Tyr 85
90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105219107PRTArtificial Sequence15G11-DI3 VL
219Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1
5 10 15Asp Arg Val Thr Ile Thr
Cys Arg Ala Ser Gln Glu Ile Ser Ser Tyr 20 25
30Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys
Leu Leu Ile 35 40 45Tyr Ala Ala
Ser Ser Leu Asp Thr Gly Val Pro Ser Arg Phe Ser Gly 50
55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
Ser Leu Gln Pro65 70 75
80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Ser Pro Tyr
85 90 95Thr Phe Gly Gly Gly Thr
Lys Val Glu Ile Lys 100 105220107PRTArtificial
Sequence15G11-DI4 VL 220Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser
Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Thr Tyr
20 25 30Leu Ser Trp Tyr Gln Gln Lys
Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40
45Tyr Ala Ala Ser Ser Leu Asp Thr Gly Val Pro Ser Arg Phe Ser
Gly 50 55 60Ser Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70
75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr
Asp Ser Ser Pro Tyr 85 90
95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100
105221107PRTArtificial Sequence15G11-DI5 VL 221Asp Ile Gln Met Thr
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5
10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln
Glu Ile Ser Ser Tyr 20 25
30Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45Tyr Ala Ala Ser Ser Leu Asp Thr
Gly Val Pro Ser Arg Phe Ser Gly 50 55
60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80Glu Asp Phe Ala Thr
Tyr Tyr Cys Leu Gln Tyr Asp Ser Thr Pro Tyr 85
90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105222107PRTArtificial Sequence15G11-DI6 VL
222Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1
5 10 15Asp Arg Val Thr Ile Thr
Cys Arg Ala Ser Gln Glu Ile Ser Thr Tyr 20 25
30Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys
Leu Leu Ile 35 40 45Tyr Ala Ala
Ser Ser Leu Asp Thr Gly Val Pro Ser Arg Phe Ser Gly 50
55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
Ser Leu Gln Pro65 70 75
80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Thr Pro Tyr
85 90 95Thr Phe Gly Gly Gly Thr
Lys Val Glu Ile Lys 100 105223107PRTArtificial
Sequence15G11-DI7 VL 223Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser
Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Ser Tyr
20 25 30Leu Ser Trp Tyr Gln Gln Lys
Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40
45Tyr Ala Ala Ser Ser Leu Asp Thr Gly Val Pro Ser Arg Phe Ser
Gly 50 55 60Ser Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70
75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr
Asp Ser Ser Pro Tyr 85 90
95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100
105224107PRTArtificial Sequence15G11-DI8 VL 224Asp Ile Gln Met Thr
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5
10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln
Glu Ile Ser Thr Tyr 20 25
30Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45Tyr Ala Ala Ser Ser Leu Asp Thr
Gly Val Pro Ser Arg Phe Ser Gly 50 55
60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80Glu Asp Phe Ala Thr
Tyr Tyr Cys Leu Gln Tyr Asp Ser Ser Pro Tyr 85
90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105225107PRTArtificial Sequence15G11-DI9 VL
225Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1
5 10 15Asp Arg Val Thr Ile Thr
Cys Arg Ala Ser Gln Glu Ile Ser Thr Tyr 20 25
30Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys
Leu Leu Ile 35 40 45Tyr Ala Ala
Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50
55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
Ser Leu Gln Pro65 70 75
80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Ser Pro Leu
85 90 95Thr Phe Gly Gly Gly Thr
Lys Val Glu Ile Lys 100 105226107PRTArtificial
Sequence15G11-DI10 VL 226Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser
Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Ser Tyr
20 25 30Leu Ser Trp Tyr Gln Gln Lys
Pro Gly Lys Ala Pro Lys Leu Leu Ala 35 40
45Tyr Ala Ala Ser Ser Leu Asp Ser Gly Val Pro Ser Arg Phe Ser
Gly 50 55 60Ser Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70
75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr
Asp Ser Thr Pro Tyr 85 90
95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100
105227107PRTArtificial Sequence15G11-DI11 VL 227Asp Ile Gln Met Thr
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5
10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln
Glu Ala Ser Ser Tyr 20 25
30Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ala
35 40 45Tyr Ala Ala Ser Ser Leu Asp Ser
Gly Val Pro Ser Arg Phe Ser Gly 50 55
60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80Glu Asp Phe Ala Thr
Tyr Tyr Cys Leu Gln Tyr Asp Ser Ser Pro Tyr 85
90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105228119PRTArtificial Sequence15G11-DI1 VH
228Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1
5 10 15Ser Leu Arg Leu Ser Cys
Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25
30Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
Glu Trp Val 35 40 45Ser Thr Ile
Ser Asp Ser Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val 50
55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys
Asn Ser Leu Tyr65 70 75
80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Ala Arg Glu Leu Gly Asp
Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100
105 110Thr Leu Val Thr Val Ser Ser
115229119PRTArtificial Sequence15G11-DI2 VH 229Gln Val Gln Leu Val Glu
Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5
10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Ser Asp Tyr 20 25 30Gly
Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35
40 45Ser Thr Ile Ser Asp Ser Gly Ser Tyr
Thr Tyr Tyr Pro Asp Ser Val 50 55
60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65
70 75 80Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95Ala Arg Glu Leu Gly Asp Tyr Asp Gly Phe Asp
Tyr Trp Gly Gln Gly 100 105
110Thr Leu Val Thr Val Ser Ser 115230119PRTArtificial
Sequence15G11-DI3 VH 230Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val
Lys Pro Gly Gly1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30Gly Met Ser Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Ser Thr Ile Ser Asp Ser Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser
Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70
75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90
95Ala Arg Glu Leu Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly
100 105 110Thr Leu Val Thr Val Ser
Ser 115231119PRTArtificial Sequence15G11-DI4 VH 231Gln Val Gln Leu
Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5
10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
Phe Thr Phe Ser Asp Tyr 20 25
30Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45Ser Thr Ile Ser Asp Ser Gly Ser
Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55
60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65
70 75 80Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95Ala Arg Glu Leu Gly Asp Tyr Asp Gly Phe Asp
Tyr Trp Gly Gln Gly 100 105
110Thr Leu Val Thr Val Ser Ser 115232119PRTArtificial
Sequence15G11-DI5 VH 232Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val
Lys Pro Gly Gly1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30Ser Met Ser Trp Ile Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Ser Thr Ile Ser Asp Ser Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser
Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70
75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90
95Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Phe Trp Gly Gln Gly
100 105 110Thr Leu Val Thr Val Ser
Ser 115233119PRTArtificial Sequence15G11-DI6 VH 233Gln Val Gln Leu
Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5
10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
Phe Thr Phe Ser Asp Tyr 20 25
30Ser Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45Ser Thr Ile Ser Asp Ser Gly Thr
Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55
60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65
70 75 80Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp
Phe Trp Gly Gln Gly 100 105
110Thr Leu Val Thr Val Ser Ser 115234119PRTArtificial
Sequence15G11-DI7 VH 234Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val
Lys Pro Gly Gly1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30Ser Met Ser Trp Ile Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Ser Thr Ile Ser Asp Ser Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser
Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70
75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90
95Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Phe Trp Gly Gln Gly
100 105 110Thr Leu Val Thr Val Ser
Ser 115235119PRTArtificial Sequence15G11-DI8 VH 235Gln Val Gln Leu
Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5
10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
Phe Thr Phe Ser Asp Tyr 20 25
30Ser Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45Ser Thr Ile Ser Asp Ser Gly Thr
Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55
60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65
70 75 80Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp
Phe Trp Gly Gln Gly 100 105
110Thr Leu Val Thr Val Ser Ser 115236119PRTArtificial
Sequence15G11-DI9 VH 236Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val
Lys Pro Gly Gly1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30Ser Met Ser Trp Ile Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Ser Thr Ile Ser Asp Ser Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser
Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70
75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90
95Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Phe Trp Gly Gln Gly
100 105 110Thr Leu Val Thr Val Ser
Ser 115237119PRTArtificial Sequence15G11-DI10 VH 237Gln Val Gln
Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5
10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser
Gly Phe Thr Phe Ser Asp Tyr 20 25
30Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45Ser Thr Ile Ser Asp Ser Gly
Ser Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55
60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65
70 75 80Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95Ala Arg Glu Leu Gly Asp Tyr Asp Gly Phe
Asp Tyr Trp Gly Gln Gly 100 105
110Thr Leu Val Thr Val Ser Ser 115238119PRTArtificial
Sequence15G11-DI11 VH 238Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val
Lys Pro Gly Gly1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30Gly Met Ser Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Ser Thr Ile Ser Asp Ser Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser
Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70
75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90
95Ala Arg Glu Leu Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly
100 105 110Thr Leu Val Thr Val Ser
Ser 115239327PRTHomo sapiens 239Ala Ser Thr Lys Gly Pro Ser Val
Phe Pro Leu Ala Pro Cys Ser Arg1 5 10
15Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys
Asp Tyr 20 25 30Phe Pro Glu
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35
40 45Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
Ser Gly Leu Tyr Ser 50 55 60Leu Ser
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr65
70 75 80Tyr Thr Cys Asn Val Asp His
Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90
95Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys
Pro Ala Pro 100 105 110Glu Phe
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 115
120 125Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
Val Thr Cys Val Val Val 130 135 140Asp
Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp145
150 155 160Gly Val Glu Val His Asn
Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe 165
170 175Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val
Leu His Gln Asp 180 185 190Trp
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu 195
200 205Pro Ser Ser Ile Glu Lys Thr Ile Ser
Lys Ala Lys Gly Gln Pro Arg 210 215
220Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys225
230 235 240Asn Gln Val Ser
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245
250 255Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
Pro Glu Asn Asn Tyr Lys 260 265
270Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
275 280 285Arg Leu Thr Val Asp Lys Ser
Arg Trp Gln Glu Gly Asn Val Phe Ser 290 295
300Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
Ser305 310 315 320Leu Ser
Leu Ser Leu Gly Lys 325240327PRTHomo sapiens 240Ala Ser
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg1 5
10 15Ser Thr Ser Glu Ser Thr Ala Ala
Leu Gly Cys Leu Val Lys Asp Tyr 20 25
30Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser 35 40 45Gly Val His Thr Phe
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55
60Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr
Lys Thr65 70 75 80Tyr
Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95Arg Val Glu Ser Lys Tyr Gly
Pro Pro Cys Pro Pro Cys Pro Ala Pro 100 105
110Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
Pro Lys 115 120 125Asp Thr Leu Met
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 130
135 140Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn
Trp Tyr Val Asp145 150 155
160Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe
165 170 175Asn Ser Thr Tyr Arg
Val Val Ser Val Leu Thr Val Leu His Gln Asp 180
185 190Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys Gly Leu 195 200 205Pro Ser
Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 210
215 220Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln
Glu Glu Met Thr Lys225 230 235
240Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
245 250 255Ile Ala Val Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 260
265 270Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
Phe Phe Leu Tyr Ser 275 280 285Arg
Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser 290
295 300Cys Ser Val Met His Glu Ala Leu His Asn
His Tyr Thr Gln Lys Ser305 310 315
320Leu Ser Leu Ser Leu Gly Lys 325241330PRTHomo
sapiens 241Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser
Lys1 5 10 15Ser Thr Ser
Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20
25 30Phe Pro Glu Pro Val Thr Val Ser Trp Asn
Ser Gly Ala Leu Thr Ser 35 40
45Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50
55 60Leu Ser Ser Val Val Thr Val Pro Ser
Ser Ser Leu Gly Thr Gln Thr65 70 75
80Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val
Asp Lys 85 90 95Lys Val
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100
105 110Pro Ala Pro Glu Leu Leu Gly Gly Pro
Ser Val Phe Leu Phe Pro Pro 115 120
125Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140Val Val Val Asp Val Ser His
Glu Asp Pro Glu Val Lys Phe Asn Trp145 150
155 160Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
Lys Pro Arg Glu 165 170
175Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190His Gln Asp Trp Leu Asn
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200
205Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala
Lys Gly 210 215 220Gln Pro Arg Glu Pro
Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu225 230
235 240Leu Thr Lys Asn Gln Val Ser Leu Thr Cys
Leu Val Lys Gly Phe Tyr 245 250
255Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
260 265 270Asn Tyr Lys Thr Thr
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275
280 285Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
Gln Gln Gly Asn 290 295 300Val Phe Ser
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr305
310 315 320Gln Lys Ser Leu Ser Leu Ser
Pro Gly Lys 325 330242330PRTHomo sapiens
242Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys1
5 10 15Ser Thr Ser Gly Gly Thr
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25
30Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala
Leu Thr Ser 35 40 45Gly Val His
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50
55 60Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
Gly Thr Gln Thr65 70 75
80Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95Lys Val Glu Pro Lys Ser
Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100
105 110Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe
Leu Phe Pro Pro 115 120 125Lys Pro
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130
135 140Val Val Val Asp Val Ser His Glu Asp Pro Glu
Val Lys Phe Asn Trp145 150 155
160Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175Glu Gln Tyr Asn
Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180
185 190His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
Cys Lys Val Ser Asn 195 200 205Lys
Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210
215 220Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
Pro Pro Ser Arg Asp Glu225 230 235
240Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
Tyr 245 250 255Pro Ser Asp
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260
265 270Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
Ser Asp Gly Ser Phe Phe 275 280
285Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290
295 300Val Phe Ser Cys Ser Val Met His
Glu Ala Leu His Asn His Tyr Thr305 310
315 320Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330243326PRTHomo sapiens 243Ala Ser Thr Lys Gly
Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg1 5
10 15Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys
Leu Val Lys Asp Tyr 20 25
30Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45Gly Val His Thr Phe Pro Ala Val
Leu Gln Ser Ser Gly Leu Tyr Ser 50 55
60Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr65
70 75 80Tyr Thr Cys Asn Val
Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85
90 95Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro
Pro Cys Pro Ala Pro 100 105
110Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
115 120 125Thr Leu Met Ile Ser Arg Thr
Pro Glu Val Thr Cys Val Val Val Asp 130 135
140Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp
Gly145 150 155 160Val Glu
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn
165 170 175Ser Thr Phe Arg Val Val Ser
Val Leu Thr Val Val His Gln Asp Trp 180 185
190Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly
Leu Pro 195 200 205Ala Pro Ile Glu
Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu 210
215 220Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu
Met Thr Lys Asn225 230 235
240Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile
245 250 255Ser Val Glu Trp Glu
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 260
265 270Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe
Leu Tyr Ser Lys 275 280 285Leu Thr
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 290
295 300Ser Val Met His Glu Ala Leu His Asn His Tyr
Thr Gln Lys Ser Leu305 310 315
320Ser Leu Ser Pro Gly Lys 325244330PRTHomo sapiens
244Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys1
5 10 15Ser Thr Ser Gly Gly Thr
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25
30Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala
Leu Thr Ser 35 40 45Gly Val His
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50
55 60Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
Gly Thr Gln Thr65 70 75
80Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95Lys Val Glu Pro Lys Ser
Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100
105 110Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
Leu Phe Pro Pro 115 120 125Lys Pro
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130
135 140Val Val Val Asp Val Ser His Glu Asp Pro Glu
Val Lys Phe Asn Trp145 150 155
160Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175Glu Gln Tyr Asn
Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180
185 190His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
Cys Lys Val Ser Asn 195 200 205Lys
Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210
215 220Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
Pro Pro Ser Arg Glu Glu225 230 235
240Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
Tyr 245 250 255Pro Ser Asp
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260
265 270Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
Ser Asp Gly Ser Phe Phe 275 280
285Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290
295 300Val Phe Ser Cys Ser Val Met His
Glu Ala Leu His Asn His Tyr Thr305 310
315 320Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330245329PRTHomo sapiens 245Ala Ser Thr Lys Gly
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys1 5
10 15Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
Leu Val Lys Asp Tyr 20 25
30Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45Gly Val His Thr Phe Pro Ala Val
Leu Gln Ser Ser Gly Leu Tyr Ser 50 55
60Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr65
70 75 80Tyr Ile Cys Asn Val
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85
90 95Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
Thr Cys Pro Pro Cys 100 105
110Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro
115 120 125Lys Pro Lys Asp Thr Leu Met
Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135
140Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn
Trp145 150 155 160Tyr Val
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175Glu Gln Tyr Asn Ser Thr Tyr
Arg Val Val Ser Val Leu Thr Val Leu 180 185
190His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
Ser Asn 195 200 205Lys Ala Leu Pro
Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210
215 220Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
Ser Arg Glu Glu225 230 235
240Met Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro
245 250 255Ser Asp Ile Ala Val
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 260
265 270Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
Ser Phe Phe Leu 275 280 285Tyr Ser
Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 290
295 300Phe Ser Cys Ser Val Met His Glu Ala Leu His
Asn His Tyr Thr Gln305 310 315
320Lys Ser Leu Ser Leu Ser Pro Gly Lys
3252461342PRTHomo sapiens 246Met Arg Ala Asn Asp Ala Leu Gln Val Leu Gly
Leu Leu Phe Ser Leu1 5 10
15Ala Arg Gly Ser Glu Val Gly Asn Ser Gln Ala Val Cys Pro Gly Thr
20 25 30Leu Asn Gly Leu Ser Val Thr
Gly Asp Ala Glu Asn Gln Tyr Gln Thr 35 40
45Leu Tyr Lys Leu Tyr Glu Arg Cys Glu Val Val Met Gly Asn Leu
Glu 50 55 60Ile Val Leu Thr Gly His
Asn Ala Asp Leu Ser Phe Leu Gln Trp Ile65 70
75 80Arg Glu Val Thr Gly Tyr Val Leu Val Ala Met
Asn Glu Phe Ser Thr 85 90
95Leu Pro Leu Pro Asn Leu Arg Val Val Arg Gly Thr Gln Val Tyr Asp
100 105 110Gly Lys Phe Ala Ile Phe
Val Met Leu Asn Tyr Asn Thr Asn Ser Ser 115 120
125His Ala Leu Arg Gln Leu Arg Leu Thr Gln Leu Thr Glu Ile
Leu Ser 130 135 140Gly Gly Val Tyr Ile
Glu Lys Asn Asp Lys Leu Cys His Met Asp Thr145 150
155 160Ile Asp Trp Arg Asp Ile Val Arg Asp Arg
Asp Ala Glu Ile Val Val 165 170
175Lys Asp Asn Gly Arg Ser Cys Pro Pro Cys His Glu Val Cys Lys Gly
180 185 190Arg Cys Trp Gly Pro
Gly Ser Glu Asp Cys Gln Thr Leu Thr Lys Thr 195
200 205Ile Cys Ala Pro Gln Cys Asn Gly His Cys Phe Gly
Pro Asn Pro Asn 210 215 220Gln Cys Cys
His Asp Glu Cys Ala Gly Gly Cys Ser Gly Pro Gln Asp225
230 235 240Thr Asp Cys Phe Ala Cys Arg
His Phe Asn Asp Ser Gly Ala Cys Val 245
250 255Pro Arg Cys Pro Gln Pro Leu Val Tyr Asn Lys Leu
Thr Phe Gln Leu 260 265 270Glu
Pro Asn Pro His Thr Lys Tyr Gln Tyr Gly Gly Val Cys Val Ala 275
280 285Ser Cys Pro His Asn Phe Val Val Asp
Gln Thr Ser Cys Val Arg Ala 290 295
300Cys Pro Pro Asp Lys Met Glu Val Asp Lys Asn Gly Leu Lys Met Cys305
310 315 320Glu Pro Cys Gly
Gly Leu Cys Pro Lys Ala Cys Glu Gly Thr Gly Ser 325
330 335Gly Ser Arg Phe Gln Thr Val Asp Ser Ser
Asn Ile Asp Gly Phe Val 340 345
350Asn Cys Thr Lys Ile Leu Gly Asn Leu Asp Phe Leu Ile Thr Gly Leu
355 360 365Asn Gly Asp Pro Trp His Lys
Ile Pro Ala Leu Asp Pro Glu Lys Leu 370 375
380Asn Val Phe Arg Thr Val Arg Glu Ile Thr Gly Tyr Leu Asn Ile
Gln385 390 395 400Ser Trp
Pro Pro His Met His Asn Phe Ser Val Phe Ser Asn Leu Thr
405 410 415Thr Ile Gly Gly Arg Ser Leu
Tyr Asn Arg Gly Phe Ser Leu Leu Ile 420 425
430Met Lys Asn Leu Asn Val Thr Ser Leu Gly Phe Arg Ser Leu
Lys Glu 435 440 445Ile Ser Ala Gly
Arg Ile Tyr Ile Ser Ala Asn Arg Gln Leu Cys Tyr 450
455 460His His Ser Leu Asn Trp Thr Lys Val Leu Arg Gly
Pro Thr Glu Glu465 470 475
480Arg Leu Asp Ile Lys His Asn Arg Pro Arg Arg Asp Cys Val Ala Glu
485 490 495Gly Lys Val Cys Asp
Pro Leu Cys Ser Ser Gly Gly Cys Trp Gly Pro 500
505 510Gly Pro Gly Gln Cys Leu Ser Cys Arg Asn Tyr Ser
Arg Gly Gly Val 515 520 525Cys Val
Thr His Cys Asn Phe Leu Asn Gly Glu Pro Arg Glu Phe Ala 530
535 540His Glu Ala Glu Cys Phe Ser Cys His Pro Glu
Cys Gln Pro Met Glu545 550 555
560Gly Thr Ala Thr Cys Asn Gly Ser Gly Ser Asp Thr Cys Ala Gln Cys
565 570 575Ala His Phe Arg
Asp Gly Pro His Cys Val Ser Ser Cys Pro His Gly 580
585 590Val Leu Gly Ala Lys Gly Pro Ile Tyr Lys Tyr
Pro Asp Val Gln Asn 595 600 605Glu
Cys Arg Pro Cys His Glu Asn Cys Thr Gln Gly Cys Lys Gly Pro 610
615 620Glu Leu Gln Asp Cys Leu Gly Gln Thr Leu
Val Leu Ile Gly Lys Thr625 630 635
640His Leu Thr Met Ala Leu Thr Val Ile Ala Gly Leu Val Val Ile
Phe 645 650 655Met Met Leu
Gly Gly Thr Phe Leu Tyr Trp Arg Gly Arg Arg Ile Gln 660
665 670Asn Lys Arg Ala Met Arg Arg Tyr Leu Glu
Arg Gly Glu Ser Ile Glu 675 680
685Pro Leu Asp Pro Ser Glu Lys Ala Asn Lys Val Leu Ala Arg Ile Phe 690
695 700Lys Glu Thr Glu Leu Arg Lys Leu
Lys Val Leu Gly Ser Gly Val Phe705 710
715 720Gly Thr Val His Lys Gly Val Trp Ile Pro Glu Gly
Glu Ser Ile Lys 725 730
735Ile Pro Val Cys Ile Lys Val Ile Glu Asp Lys Ser Gly Arg Gln Ser
740 745 750Phe Gln Ala Val Thr Asp
His Met Leu Ala Ile Gly Ser Leu Asp His 755 760
765Ala His Ile Val Arg Leu Leu Gly Leu Cys Pro Gly Ser Ser
Leu Gln 770 775 780Leu Val Thr Gln Tyr
Leu Pro Leu Gly Ser Leu Leu Asp His Val Arg785 790
795 800Gln His Arg Gly Ala Leu Gly Pro Gln Leu
Leu Leu Asn Trp Gly Val 805 810
815Gln Ile Ala Lys Gly Met Tyr Tyr Leu Glu Glu His Gly Met Val His
820 825 830Arg Asn Leu Ala Ala
Arg Asn Val Leu Leu Lys Ser Pro Ser Gln Val 835
840 845Gln Val Ala Asp Phe Gly Val Ala Asp Leu Leu Pro
Pro Asp Asp Lys 850 855 860Gln Leu Leu
Tyr Ser Glu Ala Lys Thr Pro Ile Lys Trp Met Ala Leu865
870 875 880Glu Ser Ile His Phe Gly Lys
Tyr Thr His Gln Ser Asp Val Trp Ser 885
890 895Tyr Gly Val Thr Val Trp Glu Leu Met Thr Phe Gly
Ala Glu Pro Tyr 900 905 910Ala
Gly Leu Arg Leu Ala Glu Val Pro Asp Leu Leu Glu Lys Gly Glu 915
920 925Arg Leu Ala Gln Pro Gln Ile Cys Thr
Ile Asp Val Tyr Met Val Met 930 935
940Val Lys Cys Trp Met Ile Asp Glu Asn Ile Arg Pro Thr Phe Lys Glu945
950 955 960Leu Ala Asn Glu
Phe Thr Arg Met Ala Arg Asp Pro Pro Arg Tyr Leu 965
970 975Val Ile Lys Arg Glu Ser Gly Pro Gly Ile
Ala Pro Gly Pro Glu Pro 980 985
990His Gly Leu Thr Asn Lys Lys Leu Glu Glu Val Glu Leu Glu Pro Glu
995 1000 1005Leu Asp Leu Asp Leu Asp
Leu Glu Ala Glu Glu Asp Asn Leu Ala 1010 1015
1020Thr Thr Thr Leu Gly Ser Ala Leu Ser Leu Pro Val Gly Thr
Leu 1025 1030 1035Asn Arg Pro Arg Gly
Ser Gln Ser Leu Leu Ser Pro Ser Ser Gly 1040 1045
1050Tyr Met Pro Met Asn Gln Gly Asn Leu Gly Glu Ser Cys
Gln Glu 1055 1060 1065Ser Ala Val Ser
Gly Ser Ser Glu Arg Cys Pro Arg Pro Val Ser 1070
1075 1080Leu His Pro Met Pro Arg Gly Cys Leu Ala Ser
Glu Ser Ser Glu 1085 1090 1095Gly His
Val Thr Gly Ser Glu Ala Glu Leu Gln Glu Lys Val Ser 1100
1105 1110Met Cys Arg Ser Arg Ser Arg Ser Arg Ser
Pro Arg Pro Arg Gly 1115 1120 1125Asp
Ser Ala Tyr His Ser Gln Arg His Ser Leu Leu Thr Pro Val 1130
1135 1140Thr Pro Leu Ser Pro Pro Gly Leu Glu
Glu Glu Asp Val Asn Gly 1145 1150
1155Tyr Val Met Pro Asp Thr His Leu Lys Gly Thr Pro Ser Ser Arg
1160 1165 1170Glu Gly Thr Leu Ser Ser
Val Gly Leu Ser Ser Val Leu Gly Thr 1175 1180
1185Glu Glu Glu Asp Glu Asp Glu Glu Tyr Glu Tyr Met Asn Arg
Arg 1190 1195 1200Arg Arg His Ser Pro
Pro His Pro Pro Arg Pro Ser Ser Leu Glu 1205 1210
1215Glu Leu Gly Tyr Glu Tyr Met Asp Val Gly Ser Asp Leu
Ser Ala 1220 1225 1230Ser Leu Gly Ser
Thr Gln Ser Cys Pro Leu His Pro Val Pro Ile 1235
1240 1245Met Pro Thr Ala Gly Thr Thr Pro Asp Glu Asp
Tyr Glu Tyr Met 1250 1255 1260Asn Arg
Gln Arg Asp Gly Gly Gly Pro Gly Gly Asp Tyr Ala Ala 1265
1270 1275Met Gly Ala Cys Pro Ala Ser Glu Gln Gly
Tyr Glu Glu Met Arg 1280 1285 1290Ala
Phe Gln Gly Pro Gly His Gln Ala Pro His Val His Tyr Ala 1295
1300 1305Arg Leu Lys Thr Leu Arg Ser Leu Glu
Ala Thr Asp Ser Ala Phe 1310 1315
1320Asp Asn Pro Asp Tyr Trp His Ser Arg Leu Phe Pro Lys Ala Asn
1325 1330 1335Ala Gln Arg Thr
13402471342PRTMacaca mulatta 247Met Arg Ala Asn Gly Ala Leu Gln Val Leu
Gly Leu Leu Phe Asn Leu1 5 10
15Ala Arg Gly Ser Glu Val Gly Asn Ser Gln Ala Val Cys Pro Gly Thr
20 25 30Leu Asn Gly Leu Ser Val
Thr Gly Asp Ala Glu Asn Gln Tyr Gln Thr 35 40
45Leu Tyr Lys Leu Tyr Glu Arg Cys Glu Val Val Met Gly Asn
Leu Glu 50 55 60Ile Val Leu Thr Gly
His Asn Ala Asp Leu Ser Phe Leu Gln Trp Ile65 70
75 80Arg Glu Val Thr Gly Tyr Val Leu Val Ala
Met Asn Glu Phe Ser Thr 85 90
95Leu Pro Leu Pro Asn Leu Arg Val Val Arg Gly Thr Gln Val Tyr Asp
100 105 110Gly Lys Phe Ala Ile
Phe Val Met Leu Asn Tyr Asn Thr Asn Ser Ser 115
120 125His Ala Leu Arg Gln Leu Arg Leu Thr Gln Leu Thr
Glu Ile Leu Ser 130 135 140Gly Gly Val
Tyr Ile Glu Lys Asn Asp Lys Leu Cys His Met Asp Thr145
150 155 160Ile Asp Trp Lys Asp Ile Val
Arg Asp Gln Asp Ala Glu Ile Val Val 165
170 175Lys Asp Asn Gly Arg Ser Cys Pro Leu Cys His Glu
Val Cys Lys Gly 180 185 190Arg
Cys Trp Gly Pro Gly Pro Glu Asp Cys Gln Thr Leu Thr Lys Thr 195
200 205Ile Cys Ala Pro Gln Cys Asn Gly His
Cys Phe Gly Pro Asn Pro Asn 210 215
220Gln Cys Cys His Asp Glu Cys Ala Gly Gly Cys Ser Gly Pro Gln Asp225
230 235 240Thr Asp Cys Phe
Ala Cys Arg His Phe Asn Asp Ser Gly Ala Cys Val 245
250 255Pro Arg Cys Pro Gln Pro Leu Val Tyr Asn
Lys Leu Thr Phe Gln Leu 260 265
270Glu Pro Asn Pro His Thr Lys Tyr Gln Tyr Gly Gly Val Cys Val Ala
275 280 285Ser Cys Pro His Asn Phe Val
Val Asp Gln Thr Ser Cys Val Arg Ala 290 295
300Cys Pro Pro Asp Lys Met Glu Val Asp Lys Asn Gly Leu Lys Met
Cys305 310 315 320Glu Pro
Cys Gly Gly Leu Cys Pro Lys Ala Cys Glu Gly Thr Gly Ser
325 330 335Gly Ser Arg Phe Gln Thr Val
Asp Ser Ser Asn Ile Asp Gly Phe Val 340 345
350Asn Cys Thr Lys Ile Leu Gly Asn Leu Asp Phe Leu Ile Thr
Gly Leu 355 360 365Asn Gly Asp Pro
Trp His Lys Ile Pro Ala Leu Asp Pro Glu Lys Leu 370
375 380Asn Val Phe Arg Thr Val Arg Glu Ile Thr Gly Tyr
Leu Asn Ile Gln385 390 395
400Ser Trp Pro Pro His Met Tyr Asn Phe Ser Val Phe Ser Asn Leu Thr
405 410 415Thr Ile Gly Gly Arg
Ser Leu Tyr Asn Arg Gly Phe Ser Leu Leu Ile 420
425 430Met Lys Asn Leu Asn Val Thr Ser Leu Gly Phe Arg
Ser Leu Lys Glu 435 440 445Ile Ser
Ala Gly Arg Ile Tyr Ile Ser Ala Asn Arg Gln Leu Cys Tyr 450
455 460His His Ser Leu Asn Trp Thr Lys Val Leu Arg
Gly Pro Thr Glu Glu465 470 475
480Arg Leu Asp Ile Lys His Asn Arg Pro Arg Arg Asp Cys Val Ala Glu
485 490 495Gly Lys Val Cys
Asp Pro Leu Cys Ser Ser Gly Gly Cys Trp Gly Pro 500
505 510Gly Pro Gly Gln Cys Leu Ser Cys Arg Asn Tyr
Ser Arg Gly Gly Val 515 520 525Cys
Val Thr His Cys Asn Phe Leu Asn Gly Glu Pro Arg Glu Phe Ala 530
535 540His Glu Ala Glu Cys Phe Ser Cys His Pro
Glu Cys Gln Pro Met Glu545 550 555
560Gly Thr Ala Thr Cys Asn Gly Ser Gly Ser Asp Thr Cys Ala Gln
Cys 565 570 575Ala His Phe
Arg Asp Gly Pro His Cys Val Ser Ser Cys Pro His Gly 580
585 590Val Leu Gly Ala Lys Gly Pro Ile Tyr Lys
Tyr Pro Asp Val Gln Asn 595 600
605Glu Cys Arg Pro Cys His Glu Asn Cys Thr Gln Gly Cys Lys Gly Pro 610
615 620Glu Leu Gln Asp Cys Leu Gly Gln
Thr Leu Val Leu Ile Gly Lys Thr625 630
635 640His Leu Thr Met Ala Leu Thr Val Ile Ala Gly Leu
Val Val Ile Phe 645 650
655Met Met Leu Gly Gly Thr Phe Leu Tyr Trp Arg Gly Arg Arg Ile Gln
660 665 670Asn Lys Arg Ala Met Arg
Arg Tyr Leu Glu Arg Gly Glu Ser Ile Glu 675 680
685Pro Leu Asp Pro Ser Glu Lys Ala Asn Lys Val Leu Ala Arg
Ile Phe 690 695 700Lys Glu Thr Glu Leu
Arg Lys Leu Lys Val Leu Gly Ser Gly Val Phe705 710
715 720Gly Thr Val His Lys Gly Val Trp Ile Pro
Glu Gly Glu Ser Ile Lys 725 730
735Ile Pro Val Cys Ile Lys Ile Ile Glu Asp Lys Ser Gly Arg Gln Ser
740 745 750Phe Gln Ala Val Thr
Asp His Met Leu Ala Ile Gly Ser Leu Asp His 755
760 765Ala His Ile Val Arg Leu Leu Gly Leu Cys Pro Gly
Ser Ser Leu Gln 770 775 780Leu Val Thr
Gln Tyr Leu Pro Leu Gly Ser Leu Leu Asp His Val Arg785
790 795 800Gln His Arg Gly Ala Leu Gly
Pro Gln Leu Leu Leu Asn Trp Gly Val 805
810 815Gln Ile Ala Lys Gly Met Tyr Tyr Leu Glu Glu His
Gly Met Val His 820 825 830Arg
Asn Leu Ala Ala Arg Asn Val Leu Leu Lys Ser Pro Ser Gln Val 835
840 845Gln Val Ala Asp Phe Gly Val Ala Asp
Leu Leu Pro Pro Asp Asp Lys 850 855
860Gln Leu Leu Tyr Ser Glu Ala Lys Thr Pro Ile Lys Trp Met Ala Leu865
870 875 880Glu Ser Ile His
Phe Gly Lys Tyr Thr His Gln Ser Asp Val Trp Ser 885
890 895Tyr Gly Val Thr Val Trp Glu Leu Met Thr
Phe Gly Ala Glu Pro Tyr 900 905
910Ala Gly Leu Arg Leu Ala Glu Val Pro Asp Leu Leu Glu Lys Gly Glu
915 920 925Arg Leu Ala Gln Pro Gln Ile
Cys Thr Ile Asp Val Tyr Met Val Met 930 935
940Val Lys Cys Trp Met Ile Asp Glu Asn Ile Arg Pro Thr Phe Lys
Glu945 950 955 960Leu Ala
Asn Glu Phe Thr Arg Met Ala Arg Asp Pro Pro Arg Tyr Leu
965 970 975Val Ile Lys Arg Glu Ser Gly
Pro Gly Ile Ala Pro Gly Pro Glu Pro 980 985
990His Gly Leu Thr Asn Lys Lys Leu Glu Glu Val Glu Leu Glu
Pro Glu 995 1000 1005Leu Asp Leu
Asp Leu Asp Leu Glu Ala Glu Glu Asp Asn Leu Ala 1010
1015 1020Thr Thr Thr Leu Gly Ser Ala Leu Ser Leu Pro
Val Gly Thr Leu 1025 1030 1035Asn Arg
Pro Arg Gly Ser Gln Ser Leu Leu Ser Pro Ser Ser Gly 1040
1045 1050Tyr Met Pro Met Asn Gln Gly Asn Leu Gly
Glu Ala Cys Gln Glu 1055 1060 1065Ser
Ala Val Ser Gly Ser Ser Glu Trp Cys Pro Arg Pro Val Ser 1070
1075 1080Leu His Pro Met Pro Arg Gly Cys Leu
Ala Ser Glu Ser Ser Glu 1085 1090
1095Gly His Val Thr Gly Ser Glu Ala Glu Leu Gln Glu Lys Val Ser
1100 1105 1110Thr Cys Arg Ser Arg Ser
Arg Ser Arg Ser Pro Arg Pro Arg Gly 1115 1120
1125Asp Ser Ala Tyr His Ser Gln Arg His Ser Leu Leu Thr Pro
Val 1130 1135 1140Thr Pro Leu Ser Pro
Pro Gly Leu Glu Glu Glu Asp Val Asn Gly 1145 1150
1155Tyr Val Met Pro Asp Thr His Leu Lys Gly Thr Pro Ser
Ser Arg 1160 1165 1170Glu Gly Thr Leu
Ser Ser Val Gly Leu Ser Ser Val Leu Gly Thr 1175
1180 1185Glu Glu Glu Asp Glu Asp Glu Glu Tyr Glu Tyr
Met Asn Arg Arg 1190 1195 1200Arg Arg
His Ser Pro Pro Arg Pro Pro Arg Pro Ser Ser Leu Glu 1205
1210 1215Glu Leu Gly Tyr Glu Tyr Met Asp Val Gly
Ser Asp Leu Ser Ala 1220 1225 1230Ser
Leu Gly Ser Thr Gln Ser Cys Pro Leu His Pro Val Pro Val 1235
1240 1245Met Pro Thr Ala Gly Thr Thr Pro Asp
Glu Asp Tyr Glu Tyr Met 1250 1255
1260Asn Arg Gln Arg Gly Gly Ser Gly Pro Gly Gly Asp Tyr Ala Ala
1265 1270 1275Met Gly Ala Cys Pro Ala
Ser Glu Gln Gly Tyr Glu Glu Met Arg 1280 1285
1290Ala Phe Gln Gly Pro Gly His Gln Ala Pro His Val His Tyr
Ala 1295 1300 1305His Leu Lys Thr Leu
Arg Ser Leu Glu Ala Thr Asp Ser Ala Phe 1310 1315
1320Asp Asn Pro Asp Tyr Trp His Ser Arg Leu Phe Pro Lys
Ala Asn 1325 1330 1335Ala Gln Arg Thr
13402485PRTHomo sapiens 248Arg Asp Glu Leu Thr1
52494PRTHomo sapiens 249Arg Glu Glu Met12509PRTArtificial
Sequenceepitope-ablating variant peptide 250Tyr Ser Met Ser Trp Ile Arg
Gln Ala1 52519PRTArtificial Sequenceepitope-ablating
variant peptide 251Tyr Gly Met Ser Trp Val Arg Gln Ala1
52529PRTArtificial SequenceHAF epitope 252Ile Tyr Ala Ala Ser Thr Leu Asp
Ser1 5253119PRTArtificial SequenceAntibody 15G11 heavy
chain variable (VH) region 253Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu
Val Lys Pro Gly Gly1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30Gly Met Ser Trp Ile Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Ser Thr Ile Ser Asp Ser Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser
Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70
75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90
95Ala Arg Glu Leu Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly
100 105 110Thr Leu Val Thr Val Ser
Ser 115254107PRTArtificial SequenceAntibody 15G11 light chain
variable (VL) region 254Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser
Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Ser Tyr
20 25 30Leu Ser Trp Tyr Gln Gln Lys
Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40
45Tyr Ala Ala Ser Ser Leu Asp Ser Gly Val Pro Ser Arg Phe Ser
Gly 50 55 60Ser Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70
75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr
Asp Ser Thr Pro Tyr 85 90
95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100
105255119PRTArtificial SequenceAntibody 16B09 heavy chain variable
(VH) region 255Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro
Gly Gly1 5 10 15Ser Leu
Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20
25 30Ser Met Ser Trp Ile Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40
45Ser Thr Ile Ser Asp Ser Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val 50
55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ala Lys Asn Ser Leu Tyr65 70 75
80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95Ala Arg
Glu Trp Gly Asp Tyr Asp Gly Phe Asp Phe Trp Gly Gln Gly 100
105 110Thr Leu Val Thr Val Ser Ser
115256107PRTArtificial SequenceAntibody 16B09 light chain variable (VL)
region 256Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val
Gly1 5 10 15Asp Arg Val
Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Ser Tyr 20
25 30Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys
Ala Pro Lys Leu Leu Ile 35 40
45Tyr Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50
55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Ser Leu Gln Pro65 70 75
80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Thr
Pro Leu 85 90 95Thr Phe
Gly Gly Gly Thr Lys Val Glu Ile Lys 100
10525716PRTArtificial Sequenceportion of anti-ERBB3 15G11 antibody VH
regionmisc_feature(4)..(4)Xaa is Tyr, Ala, Gly, His, Ser or any non-
hydrophobic amino acidmisc_feature(5)..(5)Xaa is Gly, Ala, Ser, Trp or
Tyrmisc_feature(6)..(6)Xaa is Met, Ala, Phe, Ser, Val, Trp or
Tyrmisc_feature(7)..(7)Xaa is Ser, Gly, Asn or Tyrmisc_feature(9)..(9)Xaa
is Ile or Valmisc_feature(12)..(12)Xaa is Ala, Pro or Thr 257Phe Ser Asp
Xaa Xaa Xaa Xaa Trp Xaa Arg Gln Xaa Pro Gly Lys Gly1 5
10 1525816PRTArtificial Sequenceportion of
anti-ERBB3 15G11 antibody VL regionmisc_feature(4)..(4)Xaa is Ile, Ala,
Gly, Ser or any non- hydrophobic amino acidmisc_feature(5)..(5)Xaa
is Ser or Aspmisc_feature(6)..(6)Xaa is Ser, Asp, Lys or
Thrmisc_feature(7)..(7)Xaa is Tyr, Ala, Arg, Ser or
Trpmisc_feature(9)..(9)Xaa is Ser, Ala, Gly, His, Asn or
Tyrmisc_feature(12)..(12)Xaa is Gln or Arg 258Ser Gln Glu Xaa Xaa Xaa Xaa
Leu Xaa Trp Tyr Xaa Gln Lys Pro Gly1 5 10
1525915PRTArtificial Sequenceportion of anti-ERBB3 15G11
antibody VL regionmisc_feature(6)..(6)Xaa is Ala or
Aspmisc_feature(7)..(7)Xaa is Ala or Aspmisc_feature(12)..(12)Xaa is Ser,
Asp, Pro or Thr 259Lys Leu Leu Ile Tyr Xaa Xaa Ser Ser Leu Asp Xaa Gly
Val Pro1 5 10
1526015PRTArtificial Sequenceportion of anti-ERBB3 15G11 antibody VL
regionmisc_feature(4)..(4)Xaa is Leu, Ala, Gly, His, Gln, Ser or any non-
hydrophobic amino acidmisc_feature(5)..(5)Xaa is Gln, Ala, His, Ser
or Thrmisc_feature(6)..(6)Xaa is Tyr, Ala, Gly, His, Ser or
Trpmisc_feature(9)..(9)Xaa is Thr, Ala, Gly, Asn, Ser or
Tyrmisc_feature(10)..(10)Xaa is Pro, Asp, Ser or Thr 260Tyr Tyr Cys Xaa
Xaa Xaa Asp Ser Xaa Xaa Tyr Thr Phe Gly Gly1 5
10 1526111PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion LCDR1 261Arg Ala Ser Gln Glu Ile
Ser Thr Tyr Leu Ser1 5
102629PRTArtificial Sequenceanti-ERBB3 antibody molecule or
antigen-binding portion LCDR3 262Leu Gln Tyr Asp Ser Ser Pro Leu
Thr1 52637PRTArtificial Sequenceanti-ERBB3 antibody
molecule or antigen-binding portion LCDR2 263Ala Ala Ser Ser Leu Asp
Thr1 5
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