Antibodies that bind cell-associated CA 125/O772P and methods of use thereof
View Patent ↗The present invention provides antibodies, and antigen-binding fragments of antibodies, fusion polypeptides and analogs that preferentially bind cell-associated CA 125/O772P polypeptides relative to shed CA 125/O772P polypeptides. The present invention further provides methods of preventing, managing, treating or ameliorating one or more symptoms associated with a CA 125/O772P-related disorder. In particular, the present invention provides methods of preventing, managing, treating, or ameliorating one or more symptoms associated with a cell proliferative disorder, such as cancer, e.g., ovarian cancer. The present invention still further provides methods for diagnosing a CA 125/O772P-related disorder or predisposition to developing such a disorder, as well as methods for identifying antibodies, and antigen-binding fragments of antibodies, that preferentially bind cell-associated CA 125/O772P polypeptides relative to shed CA 125/O772P polypeptides.
1. An isolated monoclonal antibody, or an antigen-binding antibody fragment, that binds to the amino acid sequence from residues 14-452 of SEQ ID NO:1, wherein the antibody preferentially binds cell-associated CA 125/O772P polypeptide relative to shed CA 125/O772P polypeptide.
2. The isolated antibody, or the antigen-binding antibody fragment, of claim 1 , wherein the antibody or antibody fragment, in an ELISA Competition Assay, exhibits less than 25% inhibition of binding to the peptide of SEQ ID NO:1 in the presence of a 25-fold (weight/weight) excess of shed CA 125/O772P over the peptide of SEQ ID NO:1.
3. The isolated antibody, or the antigen-binding antibody fragment, of claim 2 , wherein the antibody or antibody fragment, in an ELISA Competition Assay, exhibits less than 20% inhibition of binding to the peptide of SEQ ID NO:1 in the presence of a 25-fold (weight/weight) excess of shed CA 125/O772P over the peptide of SEQ ID NO:1.
4. The isolated antibody, or the antigen-binding antibody fragment, of claim 3 , wherein the antibody or antibody fragment, in an ELISA Competition Assay, exhibits less than 15% inhibition of binding to the peptide of SEQ ID NO:1 in the presence of a 25-fold (weight/weight) excess of shed CA 125/O772P over the peptide of SEQ ID NO:1.
5. The isolated antibody, or the antigen-binding antibody fragment, of claim 4 , wherein the antibody or antibody fragment, in an ELISA Competition Assay, exhibits less than 10% inhibition of binding to the peptide of SEQ ID NO:1 in the presence of a 25-fold (weight/weight) excess of shed CA 125/O772P over the peptide of SEQ ID NO:1.
6. The isolated antibody, or the antigen-binding antibody fragment, of claim 5 , wherein the antibody or antibody fragment, in an ELISA Competition Assay, exhibits less than 5% inhibition of binding to the peptide of SEQ ID NO:1 in the presence of a 25-fold (weight/weight) excess of shed CA 125/O772P over the peptide of SEQ ID NO:1.
7. The isolated antibody, or the antigen-binding antibody fragment, of claim 1 , wherein the antibody or antibody fragment, in a Flow Cytometry Assay, exhibits an IC 50 , as measured by percent-positive cells, of at least 0.05 mg/ml shed CA 125/O772P.
8. The isolated antibody, or the antigen-binding antibody fragment, of claim 7 , wherein the antibody or antibody fragment, in a Flow Cytometry Assay, exhibits an IC 50 , as measured by percent-positive cells, of at least 0.25 mg/ml shed CA 125/O772P.
9. The isolated antibody, or the antigen-binding antibody fragment, of claim 8 wherein the antibody or antibody fragment, in a Flow Cytometry Assay, exhibits an IC 50 , as measured by percent-positive cells, of at least 0.5 mg/ml shed CA 125/O772P.
10. The isolated antibody, or the antigen-binding antibody fragment, of claim 9 , wherein the antibody or antibody fragment, in a Flow Cytometry Assay, exhibits an IC 50 , as measured by percent-positive cells, of at least 0.75 mg/ml shed CA 125/O772P.
11. The isolated antibody, or the antigen-binding antibody fragment, of claim 10 , wherein the antibody or antibody fragment, in a Flow Cytometry Assay, exhibits an IC 50 , as measured by percent-positive cells, of at least 1.0 mg/ml shed CA 125/O772P.
12. The isolated antibody, or the antigen-binding antibody fragment, of claim 1 , wherein the antibody or antibody fragment binds the peptide of SEQ ID NO:1, but does not detectably bind shed CA 125/O772P.
13. The isolated antibody of claim 1 , wherein the antibody is an IgG class antibody.
14. The isolated antibody of claim 13 , wherein the antibody is an IgG 1 isotype.
15. The isolated antibody of claim 1 , wherein the antibody is a monoclonal antibody.
16. The isolated antibody of claim 15 , wherein the antibody is a chimeric monoclonal antibody.
17. The isolated antibody of claim 16 , wherein the chimeric monoclonal antibody comprises a Cγ1 constant region.
18. The isolated antibody of claim 16 , wherein the chimeric monoclonal antibody comprises a Cγ4 constant region.
19. The isolated antibody of claim 1 , wherein the antibody is a humanized monoclonal antibody.
20. The isolated antibody of claim 15 , wherein the antibody is a human monoclonal antibody.
21. The isolated antibody of claim 1 , wherein the antibody is a bi-specific antibody.
22. The isolated antibody of claim 1 , wherein the antibody is a multi-specific antibody.
23. The isolated antibody of claim 1 , wherein the antibody is a chimeric antibody.
24. The isolated antibody of claim 1 , wherein the antibody is a single chain antibody, a disulfide-linked F v s, a single chain F v s, or an anti-idiotypic antibody.
25. A monoclonal antibody produced by hybridoma 4E7 (ATCC® Accession No. PTA-5109), 7A11 (ATCC® Accession No. PTA-5110), 7C6 (ATCC® Accession No. PTA-5111), 7F10 (ATCC® Accession No. PTA-5112), 7G10 (ATCC® Accession No. PTA-5245), 7H1 (ATCC® Accession No. PTA-5114), 8A1 (ATCC® Accession No. PTA-5115), 8B5 (ATCC® Accession No. PTA-5116), 8C3 (ATCC® Accession No. PTA-5246), 8E3 (ATCC® Accession No. PTA-5118), 8G9 (ATCC® Accession No. PTA-5119), 15C9 (ATCC® Accession No. PTA-5106), 16C7 (ATCC® Accession No. PTA-5107), 16H9 (ATCC® Accession No. PTA-5108), 117.1 (ATCC® Accession No. PTA-4567), 325.1 (ATCC® Accession No. PTA-5120), 368.1 (ATCC® Accession No. PTA-4568), 446.1 (ATCC® Accession No. PTA-5549), 501.1 (ATCC® Accession No. PTA-4569), 621.1 (ATCC® Accession No. PTA-5121), 633.1 (ATCC® Accession No. PTA-5122), 654.1 (ATCC® Accession No. PTA-5247), 725.1 (ATCC® Accession No. PTA-5124), or 776.1 (ATCC® Accession No. PTA-4570).
26. A monoclonal antibody that competes with binding of the monoclonal antibody of claim 25 .
27. A hybridoma as deposited as hybridoma 4E7 (ATCC® Accession No. PTA-5109), 7A11 (ATCC® Accession No. PTA-5110), 7C6 (ATCC® Accession No. PTA-5111), 7F10 (ATCC® Accession No. PTA-5112), 7G10 (ATCC® Accession No. PTA-5245), 7H1 (ATCC® Accession No. PTA-5114), 8A1 (ATCC® Accession No. PTA-5115), 8B5 (ATCC® Accession No. PTA-5116), 8C3 (ATCC® Accession No. PTA-5246), 8E3 (ATCC® Accession No. PTA-5118), 8G9 (ATCC® Accession No. PTA-5119), 15C9 (ATCC® Accession No. PTA-5106), 16C7 (ATCC® Accession No. PTA-5107), 16H9 (ATCC® Accession No. PTA-5108), 117.1 (ATCC® Accession No. PTA-4567), 325.1 (ATCC® Accession No. PTA-5120), 368.1 (ATCC® Accession No. PTA-4568), 446.1 (ATCC® Accession No. PTA-5549), 501.1 (ATCC® Accession No. PTA-4569), 621.1 (ATCC® Accession No. PTA-5121), 633.1 (ATCC® Accession No. PTA-5122), 654.1 (ATCC® Accession No. PTA-5247), 725.1 (ATCC® Accession No. PTA-5124), or 776.1 (ATCC® Accession No. PTA-4570).
28. The isolated antibody, or antigen-binding antibody fragment, of claim 1 , wherein the antibody or antigen-binding antibody fragment comprises a light chain variable region that comprises the amino acid sequence depicted in SEQ ID NO:33, and wherein the antibody or antigen-binding antibody fragment comprises a heavy chain variable region that comprises the amino acid sequence depicted in SEQ ID NO:34.
29. The isolated antibody, or antigen-binding antibody fragment, of claim 1 , wherein the antibody, or antigen-binding antibody fragment binds the peptide of SEQ ID NO:1 with a K d of less than 100 nM as measured in an antigen-antibody affinity assay.
30. The isolated antibody, or antigen-binding antibody fragment, of claim 29 , wherein the antibody, or antigen-binding antibody fragment binds the peptide of SEQ ID NO:1 with a K d of less than 10 nM as measured in an antigen-antibody affinity assay.
31. The isolated antibody, or antigen-binding antibody fragment, of claim 30 , wherein the antibody, or antigen-binding antibody fragment binds the peptide of SEQ ID NO:1 with a K d of less than 1 nM as measured in an antigen-antibody affinity assay.
32. The isolated antibody, or antigen-binding antibody fragment, of claim 31 , wherein the antibody, or antigen-binding antibody fragment binds the peptide of SEQ ID NO:1 with a K d of less than 100 pM as measured in an antigen-antibody affinity assay.
33. The isolated antibody, or antigen-binding antibody fragment, of claim 32 , wherein the antibody, or antigen-binding antibody fragment binds the peptide of SEQ ID NO:1 with a K d of less than 10 pM as measured in an antigen-antibody affinity assay.
34. The isolated antibody, or the antigen-binding antibody fragment, of claim 1 , wherein the antibody or antibody fragment mediates lysis of a CA 125/O772P-positive tumor cell in an antibody-dependent cellular cytotoxicity (“ADCC”) assay.
35. The isolated antibody, or the antigen-binding antibody fragment, of claim 34 , wherein the antibody or antibody fragment mediates at least 10% lysis of CA 125/O772P-positive tumor cells in an ADCC assay at a 50:1 effector:target ratio at a concentration of 5.0 μg antibody or antigen-binding fragment per ml.
36. The isolated antibody, or the antigen-binding antibody fragment, of claim 34 , wherein the antibody or antibody fragment mediates at least 10% lysis of CA 125/O772P-positive tumor cells in an ADCC assay at a 25:1 effector:target ratio at a concentration of 5.0 μg antibody or antigen-binding fragment per ml.
37. The isolated antibody, or the antigen-binding antibody fragment, of claim 34 , wherein the antibody or antibody fragment mediates at least 10% lysis of CA 125/O772P-positive tumor cells in an ADCC assay at a 12.5:1 effector:target ratio at a concentration of 5.0 μg antibody or antigen-binding fragment per ml.
38. The isolated antibody, or the antigen-binding antibody fragment, of claim 34 , wherein the antibody or antibody fragment mediates at least 10% lysis of a CA 125/O772P-positive tumor cell in an ADCC assay at a 12.5:1 effector:target ratio at a concentration of 50 ng antibody or antigen-binding fragment per ml.
39. The isolated antibody, or the antigen-binding antibody fragment, of claim 1 , wherein the antibody or antibody fragment mediates lysis of a CA 125/O772P-positive tumor cell in an complement-dependent cytotoxity (CDC) assay.
40. The isolated antibody, or the antigen-binding antibody fragment, of claim 39 , wherein the antibody or antibody fragment mediates in a range from about 15% lysis at 5 μg/ml antibody or antigen-binding antibody fragment to about 95% lysis at about 0.1 μg/ml antibody or antigen-binding antibody fragment.
41. A fusion polypeptide comprising an antibody, or an antigen-binding antibody fragment, which preferentially binds cell-associated CA 125/O772P relative to shed CA 125/O772P, wherein the antibody, or the antigen-binding antibody fragment, binds to the amino acid sequence from residues 14-452 of SEQ ID NO:1, operably linked to a heterologous agent.
42. A hybridoma that can secrete an antibody of claim 1 .
43. The isolated antibody or antigen-binding fragment of claim 1 conjugated to a cytotoxic agent.
44. The isolated antibody or antigen-binding fragment of claim 43 , wherein the cytotoxic agent is a radioisotope.
45. The isolated antibody or antigen-binding fragment of claim 44 , wherein the radioisotope is selected from the group consisting of 125 I, 131 I, 111 In, 99m Tc and 90 Y.
46. The monoclonal antibody of claim 25 or 26 , which is conjugated to a cytotoxic agent.
47. The monoclonal antibody of claim 46 , wherein the cytotoxic agent is a radioisotope.
48. The monoclonal antibody of claim 47 , wherein the radioisotope is selected from the group consisting of 125 I, 131 I, 111 In, 99m Tc and 90 Y.
49. A monoclonal antibody selected from the group consisting of 325.1, 621.1, 633.1, 654.1, 725.1, 8G9, 7F10, 8A1, 8C3, 15C9, 8E3, 8B5, 7G10, 16C7, 7C6, 7H1, 16H9, 7A11, 4E7, 117.1, 368.1, 446.1, 501.1, and 776.1, or an antigen-binding antibody fragment thereof.
50. An isolated antibody or antigen-binding antibody fragment that competes with binding of a monoclonal antibody of claim 49 .