Anti-Laminin4 antibodies specific for LG4-5
The invention provides antibodies that specifically bind to the LG4-5 modules of the G domain of laminin α4. The antibodies can preferentially stain cancer or tumor cells or tissue. The antibodies can be used for detecting cancer, evaluating the efficacy of a cancer therapy, treating cancer, and treating obesity or obesity-related diseases, among other applications.
1. A monoclonal antibody that specifically binds to an epitope within the LG4-5 modules of the G domain of laminin α4, wherein the antibody comprises three heavy chain CDRs and three light chain CDRs of
(a) an antibody characterized by a mature heavy chain variable region of SEQ ID NO:16 and mature light chain variable region of SEQ ID NO:17;
(b) an antibody characterized by a mature heavy chain variable region of SEQ ID NO:26 and mature light chain variable region of SEQ ID NO:27;
(c) an antibody characterized by a mature heavy chain variable region of SEQ ID NO:36 and mature light chain variable region of SEQ ID NO:38; or
(d) an antibody characterized by a mature heavy chain variable region of SEQ ID NO:37 and mature light chain variable region of SEQ ID NO:38.
2. The antibody of claim 1 that is chimeric, humanized, or veneered.
3. The antibody of claim 2 that has human IgG1 kappa isotype.
4. The antibody of claim 2 that is a humanized or chimeric 15F7 antibody that specifically binds to laminin α4, wherein 15F7 is a mouse antibody characterized by a mature heavy chain variable region of SEQ ID NO:16 and a mature light chain variable region of SEQ ID NO:17.
5. The humanized antibody of claim 2 comprising a humanized heavy chain comprising three CDRs of the 15F7 heavy chain variable region (SEQ ID NO:16) and a humanized light chain comprising three CDRs of the 15F7 light chain variable region (SEQ ID NO:17).
6. The humanized antibody of claim 5 comprising a humanized mature heavy chain variable region having an amino acid sequence at least 90% identical to SEQ ID NO:58 and a humanized mature light chain variable region having an amino acid sequence at least 90% identical to SEQ ID NO:59.
7. The humanized antibody of claim 1 , provided at least one of positions L42, L49, and L69 is occupied by N, S, and K, respectively, and at least one of positions H1, H20, H27, H30, H38, H40, H48, H66, H67, H75, H82A, and H91 is occupied by E, L, Y, T, K, R, I, K, A, S, R, and F, respectively.
8. The humanized antibody of claim 7 , provided positions L42, L49, and L69 are occupied by N, S, and K, respectively, and H1, H20, H27, H30, H38, H40, H48, H66, H67, H75, H82A, and H91 are occupied by E, L, Y, T, K, R, I, K, A, S, R, and F, respectively.
9. The humanized antibody of claim 7 or 8 , provided at least one of positions L8, L45, and L80 is occupied by S, R, and T, respectively.
10. The humanized antibody of claim 7 or 8 , provided at least one of positions H41 and H69 is occupied by A and L, respectively.
11. The humanized antibody of claim 9 , provided positions L8, L45, and L80 are occupied by S, R, and T, respectively.
12. The humanized antibody of claim 10 , provided positions H41 and H69 are occupied by A and L, respectively.
13. The humanized antibody of claim 6 , comprising a mature heavy chain variable region having an amino acid sequence at least 95% identical to SEQ ID NO:58 and a mature light chain variable region having an amino acid sequence at least 95% identical to SEQ ID NO:59.
14. The humanized antibody of claim 6 , wherein the mature heavy chain variable region has an amino acid sequence of SEQ ID NO:57 and the mature light chain variable region has an amino acid sequence of SEQ ID NO:59.
15. The humanized antibody of claim 6 , wherein the mature heavy chain variable region has an amino acid sequence of SEQ ID NO:57 and the mature light chain variable region has an amino acid sequence of SEQ ID NO:60.
16. The humanized antibody of claim 6 , wherein the mature heavy chain variable region has an amino acid sequence of SEQ ID NO:58 and the mature light chain variable region has an amino acid sequence of SEQ ID NO:59.
17. The humanized antibody of claim 6 , wherein the mature heavy chain variable region has an amino acid sequence of SEQ ID NO:58 and the mature light chain variable region has an amino acid sequence of SEQ ID NO:60.
18. The antibody of claim 6 that is an intact antibody.
19. The antibody of claim 6 that is a single-chain antibody, Fab, or Fab′2 fragment.
20. The humanized antibody of claim 6 , wherein the mature light chain variable region is fused to a light chain constant region and the mature heavy chain variable region is fused to a heavy chain constant region.
21. The humanized antibody of claim 20 , wherein the heavy chain constant region is a mutant form of a natural human heavy chain constant region which has reduced binding to a Fcγ receptor relative to the natural human heavy chain constant region.
22. The humanized antibody of claim 20 , wherein the heavy chain constant region is of IgG1 isotype.
23. The humanized antibody of claim 20 , wherein the mature heavy chain variable region is fused to a heavy chain constant region having the sequence of SEQ ID NO:61 and/or the mature light chain variable region is fused to a light chain constant region having the sequence of SEQ ID NO:62.
24. The humanized antibody of claim 6 , provided any differences in CDRs of the mature heavy chain variable region and mature light chain variable region from SEQ ID NOS:16 and 17, respectively reside in positions H60-H65.
25. The antibody of claim 6 , wherein the antibody is conjugated to a therapeutic or cytotoxic agent.
26. The antibody of claim 25 , wherein the cytotoxic agent is saporin.
27. The antibody of claim 25 , wherein the cytotoxic agent is a radioisotope.
28. A pharmaceutical composition comprising the antibody of claim 1 and a pharmaceutically acceptable carrier.
29. The humanized antibody of claim 7 or 8 , provided at least one of positions L8, L45, L80, and L104 is occupied by S, R, T, and V, respectively.
30. The humanized antibody of claim 29 , provided position L104 is occupied by V.
31. The humanized antibody of claim 20 , wherein the mature heavy chain variable region is fused to a heavy chain constant region having the sequence of SEQ ID NO:61, 89, or 101 and/or the mature light chain variable region is fused to a light chain constant region having the sequence of SEQ ID NO:62 or 90.
32. The humanized antibody of claim 7 or 8 , provided at least one of positions L8, L45, L80, and L104 is occupied by S, R, T, and L, respectively.
33. The humanized antibody of claim 7 or 8 , provided at least one of positions L8, L45, L80, and L104 is occupied by P, K, P, and V, respectively.
34. The humanized antibody of claim 7 or 8 , provided at least one of positions H41 and H69 is occupied by P and I, respectively.
35. The humanized antibody of claim 32 , provided positions L8, L45, L80, and L104 are occupied by S, R, T, and L, respectively.
36. The humanized antibody of claim 34 , provided positions H41 and H69 are occupied by P and I, respectively.
37. The humanized antibody of claim 35 , provided positions H41 and H69 are occupied by P and I, respectively.