Fully human antibodies and fragments recognizing human c-Met
Human antibody fragments against c-Met have been identified through phage display technology. The high affinity and low immunogenicity make them very useful for both in vivo and in vitro applications. These novel human antibodies can greatly help the study of drug resistance in EGFR targeted therapies, and improve the diagnosis and treatment of cancer patients.
1. An isolated human anti-c-Met antibody comprising a human heavy chain variable domain and a human light chain variable domain, wherein the anti-c-Met antibody is selected from a group consisting of:
an anti-c-Met antibody comprising a human heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1 and a human light chain variable domain comprising the amino acid sequence of SEQ ID NO:9;
an anti-c-Met antibody comprising a human heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:2 and a human light chain variable domain comprising the amino acid sequence of SEQ ID NO:10;
an anti-c-Met antibody comprising a human heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:3 and a human light chain variable domain comprising the amino acid sequence of SEQ ID NO:11;
an anti-c-Met antibody comprising a human heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:4 and a human light chain variable domain comprising the amino acid sequence of SEQ ID NO:12;
an anti-c-Met antibody comprising a human heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:5 and a human light chain variable domain comprising the amino acid sequence of SEQ ID NO:13;
an anti-c-Met antibody comprising a human heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:6 and a human light chain variable domain comprising the amino acid sequence of SEQ ID NO:14;
an anti-c-Met antibody comprising a human heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:7 and a human light chain variable domain comprising the amino acid sequence of SEQ ID NO:15; and
an anti-c-Met antibody comprising a human heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:8 and a human light chain variable domain comprising the amino acid sequence of SEQ ID NO:16.
2. The isolated human anti-c-Met antibody of claim 1 , wherein the antibody or antigen binding fragment thereof is a diabody.
3. The isolated human anti-c-Met antibody of claim 2 , wherein the diabody is a cys-diabody.
4. The isolated human anti-c-Met antibody of claim 3 , wherein the cys-diabody comprises the amino acid sequence SEQ ID NOS:17, 18, 19, 20, 21, 22, 23, or 24.
5. The isolated human anti-c-Met antibody of claim 1 , wherein the antibody is a single-chain variable fragment (ScFv).
6. The isolated human anti-c-Met antibody of claim 5 , wherein the ScFv comprises the amino acid sequence SEQ ID NOS:25, 26, 27, 28, 29, 30, 31, or 32.
7. The isolated human anti-c-Met antibody of claim 1 , wherein the antibody or antigen binding fragment thereof is a minibody or a triabody.
8. The isolated human anti-c-Met antibody of claim 1 , wherein the antibody or antigen binding fragment thereof is a fully human antibody.
9. The isolated human anti-c-Met antibody of claim 1 , wherein the antibody or antigen binding fragment thereof is chimeric.
10. The isolated human anti-c-Met antibody of claim 1 , wherein the antibody or antigen binding fragment thereof is conjugated to a cytotoxic agent.
11. The isolated human anti-c-Met antibody of claim 10 , wherein the cytotoxic agent is a chemotherapeutic agent.
12. The isolated human anti-c-Met antibody of claim 11 , wherein the chemotherapeutic agent is selected from the group consisting of gemcitabine, carboplatin, taxol, and paclitaxel.
13. The isolated human anti-c-Met antibody of claim 1 , wherein the antibody or antigen binding fragment thereof is conjugated to a fluorescent molecule.
14. The isolated human anti-c-Met antibody of claim 1 , wherein the antibody or antigen binding fragment thereof is radiolabeled.
15. The isolated human anti-c-Met antibody of claim 14 , wherein the radiolabel is an iodine radiolabel.
16. The isolated human anti-c-Met antibody of claim 1 , wherein the antibody or antigen binding fragment thereof has an affinity between 0.3 nM and 9 nM to c-Met.
17. A pharmaceutical composition comprising the antibody isolated human anti-c-Met antibody of claim 1 and a physiologically acceptable carrier.
18. The pharmaceutical composition of claim 17 , wherein the composition is administered by parenteral, subcutaneous, intraperitoneal, intrapulmonary, or intranasal administration.
19. The pharmaceutical composition of claim 18 , wherein the parenteral administration comprises intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
20. The pharmaceutical composition of claim 17 , wherein the composition is co-administered with a cytotoxic agent.
21. A method of diagnosing a c-Met expressing cancer in a subject comprising administering an antibody conjugate to the subject and detecting an overexpression of c-Met in the subject,
wherein the antibody conjugate comprises the antibody of claim 1 and a detectable label,
wherein overexpression of c-Met in the subject is indicative of the presence of the cancer .
22. A method of treating a c-Met expressing patient for cancer, the method comprising administering to the patient an effective amount of the anti-c-Met antibody of claim 1 .
23. The method of claim 22 , further comprising administering to the patient an effective amount of at least one additional anti-cancer agent.
24. The method of claim 22 , wherein the at least one additional anti-cancer agent is selected from the group consisting of platinum-based chemotherapy drugs, taxanes, tyrosine kinase inhibitors, anti-EGFR antibodies, anti-ErbB2 antibodies, and combinations thereof.
25. The method of claim 22 , wherein the patient is human or mammal.
26. The isolated antibody of claim 1 , wherein the anti-cMet antibody has a 6-histidine tag.
27. The isolated antibody of claim 26 , wherein the 6-histidine tag is enzymatically cleaved off after purification of the antibody.
28. The isolated antibody of claim 1 , wherein the anti-cMet antibody has a Myc tag.
29. The isolated antibody of claim 28 , wherein the Myc tag is enzymatically cleaved off after purification of the antibody.
30. The isolated antibody of claim 1 , wherein the anti-cMet antibody has a Myc tag and a 6-Histidine tag.
31. The isolated antibody of claim 30 , wherein the Myc tag and 6-Histidine tag is enzymatically cleaved off after purification of the antibody.
32. The isolated antibody of claim 30 , wherein the anti-cMet antibody has a linker, a Myc tag, and a 6-Histidine tag,
wherein the sequence of the linker, the Myc tag, and the 6-Histidine tag comprises SEQ ID NO: 81.
33. The anti-c-Met diabody of claim 2 wherein the heavy chain variable domain sequence and light chain variable domain sequence are linked by a linker comprising SEQ ID NO: 82.
34. The anti-c-Met scFv antibody of claim 5 wherein the heavy chain variable domain sequence and light chain variable domain sequence are linked by a linker comprising SEQ ID NO: 81.
35. The isolated antibody of claim 1 , wherein the anti-cMet antibody has one or more terminal cysteines.
36. The isolated antibody of claim 35 , wherein one or more terminal cysteines is labeled with a detectable molecule comprising a fluorophore, a radio label, a luminescent label, a chemoluminescent label, or a spin label.
37. The method of claim 21 , wherein the cancer is lung cancer.
38. The method of claim 37 wherein the lung cancer is non-small cell lung cancer.