Monoclonal antibodies against stromal derived factor-1 (SDF-1)
The present disclosure provides isolated monoclonal antibodies, particularly human monoclonal antibodies, that specifically bind to SDF-1 with high affinity. Nucleic acid molecules encoding SDF-1 antibodies, expression vectors, host cells and methods for expressing the SDF-1 antibodies are also provided. Immunoconjugates, bispecific molecules and pharmaceutical compositions comprising the SDF-1 antibodies are also provided. Methods for detecting SDF-1, as well as methods for treating various B cell malignancies, including breast cancer, multiple myeloma and non-Hodgkin's lymphoma, and autoimmune disorders are disclosed.
1. A monoclonal antibody, or an antigen binding portion thereof comprising:
(a) a heavy chain variable region comprising amino acids having a sequence selected from the group consisting of SEQ ID NOs: 1, 2, 3 and 4; and
(b) a light chain variable region comprising amino acids having a sequence selected from the group consisting of SEQ ID NOs: 5, 6, 7 and 8;
wherein the antibody specifically binds to SDF-1.
2. The monoclonal antibody of claim 1 , which comprises:
(a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1; and
(b) a light chain variable region comprising the amino acid sequence of SEQ ID NO:5.
3. The monoclonal antibody of claim 1 , which comprises:
(a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2; and
(b) a light chain variable region comprising the amino acid sequence of SEQ ID NO:6.
4. The monoclonal antibody of claim 1 , which comprises:
(a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:3; and
(b) a light chain variable region comprising the amino acid sequence of SEQ ID NO:7.
5. The monoclonal antibody of claim 1 , which comprises:
(a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:4; and
(b) a light chain variable region comprising the amino acid sequence of SEQ ID NO:8.
6. A monoclonal antibody, or antigen binding portion thereof, which cross-competes for binding to SDF-1 with a reference antibody, and wherein the monoclonal antibody:
(a) binds to human SDF-1 with a KD of 1×10 −7 M or less; and
(b) binds to human SDF-1 as detectable by an immunoprecipitation assay, and wherein the reference antibody comprises:
(i) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 1 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 5;
(ii) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 2 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 6;
(iii) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 3 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 7; or
(iv) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 4 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 8.
7. The monoclonal antibody, or antigen-binding portion thereof, of claim 6 , which is a human antibody.
8. The monoclonal antibody, or antigen-binding portion thereof, of claim 6 , which is a chimeric or humanized antibody.
9. The monoclonal antibody, or antigen-binding portion thereof, of claim 7 , which is an antibody of an IgG1, IgG2 or IgG4 isotype or an antigen-binding portion thereof.
10. The monoclonal antibody, or antigen-binding portion thereof, of claim 7 , which is an antibody fragment or a single chain antibody.
11. The monoclonal antibody, or antigen-binding portion thereof, of claim 7 , wherein said antibody binds to human SDF-1 with a K D of 1×10 −8 M or less.
12. The monoclonal antibody, or antigen-binding portion thereof, of claim 7 , wherein said antibody binds to human SDF-1 with a K D of 1×10 −9 M or less.
13. The monoclonal antibody, or antigen-binding portion thereof, of claim 7 , wherein said antibody has a melting temperature of 57° C. or higher.
14. The monoclonal antibody, or antigen-binding portion thereof, of claim 7 , wherein said antibody has a melting temperature of 68° C. or higher.
15. The monoclonal antibody, or antigen-binding portion thereof, of claim 7 , wherein said antibody blocks the binding of SDF-1 to CEM cells.
16. The monoclonal antibody, or antigen-binding portion thereof, of claim 7 , wherein said antibody blocks SDF-1 induced calcium flux in CEM cells.
17. The monoclonal antibody, or antigen-binding portion thereof, of claim 7 , wherein said antibody blocks SDF-1 induced migration of CEM cells.
18. The monoclonal antibody, or antigen-binding portion thereof, of claim 7 , wherein said antibody blocks capillary tube formation in HuVEC cells.
19. The monoclonal antibody, or antigen-binding portion thereof, of claim 7 , wherein human SDF-1 comprises a polypeptide having an amino acid sequence as set forth in SEQ ID NO:44.
20. The monoclonal antibody of claim 6 , which is selected from the group consisting of:
(i) an antibody comprising:
(a) a heavy chain variable region CDR1 comprising SEQ ID NO:9;
(b) a heavy chain variable region CDR2 comprising SEQ ID NO:13;
(c) a heavy chain variable region CDR3 comprising SEQ ID NO:17;
(d) a light chain variable region CDR1 comprising SEQ ID NO:21;
(e) a light chain variable region CDR2 comprising SEQ ID NO:25; and
(f) a light chain variable region CDR3 comprising SEQ ID NO:29,
(ii) an antibody comprising:
(a) a heavy chain variable region CDR1 comprising SEQ ID NO:10;
(b) a heavy chain variable region CDR2 comprising SEQ ID NO:14;
(c) a heavy chain variable region CDR3 comprising SEQ ID NO:18;
(d) a light chain variable region CDR1 comprising SEQ ID NO:22;
(e) a light chain variable region CDR2 comprising SEQ ID NO:26; and
(f) a light chain variable region CDR3 comprising SEQ ID NO:30,
(iii) an antibody comprising:
(a) a heavy chain variable region CDR1 comprising SEQ ID NO:11;
(b) a heavy chain variable region CDR2 comprising SEQ ID NO:15;
(c) a heavy chain variable region CDR3 comprising SEQ ID NO:19;
(d) a light chain variable region CDR1 comprising SEQ ID NO:23;
(e) a light chain variable region CDR2 comprising SEQ ID NO:27; and
(f) a light chain variable region CDR3 comprising SEQ ID NO:31, and
(iv) an antibody comprising:
(a) a heavy chain variable region CDR1 comprising SEQ ID NO:12;
(b) a heavy chain variable region CDR2 comprising SEQ ID NO:16;
(c) a heavy chain variable region CDR3 comprising SEQ ID NO:20;
(d) a light chain variable region CDR1 comprising SEQ ID NO:24;
(e) a light chain variable region CDR2 comprising SEQ ID NO:28; and
(f) a light chain variable region CDR3 comprising SEQ ID NO:32.
21. A composition comprising the antibody, or antigen-binding portion thereof, of claim 6 , and a pharmaceutically acceptable carrier.
22. An immunoconjugate comprising the antibody, or antigen-binding portion thereof, of claim 6 , linked to a therapeutic agent.
23. A composition comprising the immunoconjugate of claim 22 and a pharmaceutically acceptable carrier.
24. The immunoconjugate of claim 22 , wherein the therapeutic agent is a cytotoxin.
25. A composition comprising the immunoconjugate of claim 24 and a pharmaceutically acceptable carrier.
26. The immunoconjugate of claim 22 , wherein the therapeutic agent is a radioactive isotope.
27. A composition comprising the immunoconjugate of claim 26 and a pharmaceutically acceptable carrier.
28. The monoclonal antibody of claim 6 , wherein the monoclonal antibody binds to both a monomeric and a dimeric form of SDF-1.
29. The monoclonal antibody of claim 6 , wherein the monoclonal antibody binds to a receptor binding site on the SDF-1 protein.
30. The monoclonal antibody of claim 29 , wherein the monoclonal antibody binds to amino acid residues 7-19 of the SDF-1 protein.
31. The monoclonal antibody of claim 6 , wherein the monoclonal antibody binds to amino acid residues 37-50 of the SDF-1 protein.
32. The monoclonal antibody of claim 6 , wherein the monoclonal antibody binds to the dimeric but not monomeric form of SDF-1.
33. The monoclonal antibody of claim 32 , wherein the monoclonal antibody binds to a heparin binding site of SDF-1.
34. The monoclonal antibody of claim 32 , wherein the monoclonal antibody binds to a dimerization interface of SDF-1.
35. The monoclonal antibody of claim 32 , wherein the monoclonal antibody binds to amino acid residues 24-30 of SDF-1.
36. A monoclonal antibody, or an antigen-binding portion thereof, comprising a heavy chain variable region and a light chain variable region, wherein said heavy chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 41 or 42, and wherein the antibody or antigen-binding portion thereof specifically binds to SDF-1.
37. A monoclonal antibody, or an antigen-binding portion thereof, comprising a light chain variable region and a heavy chain variable region, wherein said light chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 43, and wherein the antibody or antigen-binding portion thereof specifically binds to SDF-1.
38. The monoclonal antibody or antigen-binding portion thereof of claim 37 , wherein said heavy chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 41 or 42.