ANTI-NEUBLASTIN ANTIBODIES AND USES THEREOF

• US 20110135648A1
• Filed: 08/05/2008
• Published: 06/09/2011
• Est. Priority Date: 08/08/2007
• Status: Abandoned Application

• Current Assignee: Sinfonia Biotherapeutics AB
• Original Assignee: Biogen Idec MA Inc. (Biogen, Inc.)
• Inventors: Rossomando, Anthony, Jin, Ping
• Application Number:
  US12/672,213
• Publication Number:
  US 20110135648A1
• Time in Patent Office: Days
• US Class Current: 424/139.1
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/54 F(ab')2
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: G01N 33/6827 Total protein determination, e.g. albumin in urine

Abstract

Antibodies and antigen binding fragments that bind to neublastin polypeptides are disclosed. Also disclosed are methods of using the antibodies and antigen binding fragments in assays for detecting the presence or amount of endogenous and/or exogenous neublastin in a sample and in methods of antagonizing neublastin bioactivity.

Claims

1. An isolated antibody or antigen-binding fragment thereof that selectively binds to the polypeptide of SEQ ID NO:

1 on the same epitope as the antibody produced by the hybridoma deposited in the ATCC under Accession No. PTA-7624 or PTA-7625.

3. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody or antigen-binding fragment thereof is a monoclonal antibody, a polyclonal antibody, a humanized antibody, a fully human antibody, a single chain antibody, a chimeric antibody, an Fab fragment, an F(ab′

)2 fragment, an Fab′

fragment, an Fsc fragment, or an Fv fragment.

5. A conjugate comprising the antibody or antigen-binding fragment thereof of claim 1 linked to a detectable label.

6. The conjugate of claim 5, wherein the detectable label is horseradish peroxidase or alkaline phosphatase.

7. The conjugate of claim 5, wherein the detectable label is a fluorescent or radio-labeled marker.

8. A conjugate comprising the antibody or antigen-binding fragment thereof of claim 1 linked to a solid-phase matrix.

9. The conjugate of claim 8, wherein the solid-phase matrix is a multi-well assay plate, sepharose, agarose, or a magnetic bead.

10. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of claim 1 and a pharmaceutically acceptable carrier.

11. An isolated cell that produces the antibody or antigen-binding fragment thereof of claim 1.

13. A method of determining the presence or amount of a neublastin polypeptide in a sample, the method comprising:

contacting a sample with the antibody or antigen-binding fragment thereof of claim 1 for a time sufficient for the antibody or antigen-binding fragment thereof to bind to a neublastin polypeptide, if present in the sample; and

detecting the presence or amount of the antibody or antigen-binding fragment thereof bound to the neublastin polypeptide to thereby determine the presence or amount of the neublastin polypeptide in the sample.

14. The method of claim 13, wherein the method comprises:

prior to the detecting step, contacting the sample with a second antibody or antigen-binding fragment thereof that selectively binds to the polypeptide of SEQ ID NO;

1 for a time sufficient for the second antibody or antigen-binding fragment thereof to bind to the neublastin polypeptide bound to the antibody or antigen-binding fragment thereof of any of claims 1 to 4; and

detecting the presence or amount of the second antibody or antigen-binding fragment thereof bound to the neublastin polypeptide.

21. The method of claim 13, wherein the method is a sandwich-based immunoassay.

22. The method of claim 13, wherein the sample is obtained from a mammal.

23. The method of claim 22, wherein the neublastin polypeptide is endogenous, wild-type neublastin.

24. The method of claim 13, wherein the neublastin polypeptide comprises an amino acid sequence that is at least 80% identical to amino acids 15-113 of SEQ ID NO:

1 and the polypeptide, when dimerized, binds to a complex containing GFRalpha3 and RET, and wherein the sample is obtained from a mammal to which the neublastin polypeptide has been administered.

25. The method of claim 24, wherein the amino acid sequence is at least 90% identical to amino acids 15-113 of SEQ ID NO:

1.

26. The method of claim 24, wherein the amino acid sequence is at least 95% identical to amino acids 15-113 of SEQ ID NO:

1.

27. The method of claim 24, wherein the amino acid sequence is at least 98% identical to amino acids 15-113 of SEQ ID NO:

1.

28. The method of claim 24, wherein the polypeptide comprises amino acids 15-113 of SEQ ID NO:

1, amino acids 15-113 of SEQ ID NO;

2, amino acids 15-113 of SEQ ID NO;

3, amino acids 15-113 of SEQ ID NO;

4, amino acids 15-113 of SEQ ID NO;

5, amino acids 15-113 of SEQ ID NO;

8, or amino acids 15-113 of SEQ ID NO;

9.

29. The method of claim 24, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:

1, the amino acid sequence of SEQ ID NO;

2, the amino acid sequence of SEQ ID NO;

3, the amino acid sequence of SEQ ID NO;

4, the amino acid sequence of SEQ ID NO;

5, the amino acid sequence of SEQ ID NO;

8, or the amino acid sequence of SEQ ID NO;

9.

30. The method of claim 24, wherein the polypeptide comprises amino acids 10-113 of SEQ ID NO:

1.

31. The method of claim 13, wherein the sample is blood, serum, saliva, semen, urine, lacrimal fluid, or cerebral spinal fluid.

32. A method of antagonizing neublastin activity, the method comprising administering to a mammal an amount of the antibody or antigen-binding fragment thereof of claim 1 effective to reduce or eliminate the ability of endogenous, wild-type neublastin to bind to a complex comprising RET and GFRalpha3 and induce dimerization and autophosphorylation of RET.

33. A method of treating a cancer, the method comprising administering to a mammal having a cancer a pharmaceutical composition comprising a therapeutically effective amount of the antibody or antigen-binding fragment thereof of claim 1.

34. The method of claim 33, wherein the cancer is a pancreatic cancer.

35. The method of claim 22, wherein the mammal is a human.

2. An isolated antibody or antigen-binding fragment thereof that selectively binds to the polypeptide of SEQ ID NO:

1 and crossblocks binding of the antibody produced by the hybridoma deposited in the ATCC under Accession No. PTA-7624 or PTA-7625.

4. An antibody produced by the hybridoma deposited in the ATCC under Accession No. PTA-7624 or PTA-7625.

12. The hybridoma deposited in the ATCC under Accession No. PTA-7624 or PTA 7625.

15. A method of determining the presence or amount of a neublastin polypeptide in a sample, the method comprising:

contacting a sample with a first antibody or antigen-binding fragment thereof that selectively binds to the polypeptide of SEQ ID NO;

1 on the same epitope as the antibody produced by the hybridoma deposited in the ATCC under Accession No. PTA-7625 for a time sufficient for the first antibody or antigen-binding fragment thereof to bind to a neublastin polypeptide, if present in the sample;

contacting the sample with a second antibody or antigen-binding fragment thereof that selectively binds to the polypeptide of SEQ ID NO;

1 on the same epitope as the antibody produced by the hybridoma deposited in the ATCC under Accession No. PTA 7624 for a time sufficient for the second antibody or antigen-binding fragment thereof to bind the neublastin polypeptide, if bound to the first antibody or antigen-binding fragment thereof; and

detecting the presence or amount of the second antibody or antigen-binding fragment thereof bound to the neublastin polypeptide, to thereby determine the presence or amount of the neublastin polypeptide in the sample.

16. The method of claim 15, wherein the first antibody is produced by the hybridoma deposited in the ATCC under Accession No. PTA-7625.

17. The method of claim 15, wherein the second antibody is produced by the hybridoma deposited in the ATCC under Accession No. PTA-7624.

18. A method of determining the presence or amount of a neublastin polypeptide in a sample, the method comprising:

contacting a sample with a first antibody or antigen-binding fragment thereof that selectively binds to the polypeptide of SEQ ID NO;

1 on the same epitope as the antibody produced by the hybridoma deposited in the ATCC under Accession No. PTA-7625 for a time sufficient for the first antibody or antigen-binding fragment thereof to bind to a neublastin polypeptide, if present in the sample;

contacting the sample with a second antibody or antigen-binding fragment thereof that selectively binds to the polypeptide of SEQ ID NO;

1 on the same epitope as the antibody produced by the hybridoma deposited in the ATCC under Accession No. PTA 7625 for a time sufficient for the second antibody or antigen-binding fragment thereof to bind to the neublastin polypeptide, if bound to the first antibody or antigen-binding fragment thereof; and

detecting the presence or amount of the second antibody or antigen-binding fragment thereof bound to the neublastin polypeptide, to thereby determine the presence or amount of the neublastin polypeptide in the sample.

19. The method of claim 18, wherein the first antibody is produced by the hybridoma deposited in the ATCC under Accession No. PTA-7625.

20. The method of claim 18, wherein the second antibody is produced by the hybridoma deposited in the ATCC under Accession No. PTA-7625.