Gdf8 antibodies and use thereof

FIELD: chemistry, pharmaceutics.

SUBSTANCE: present invention refers to biotechnology and immunology. There are offered: a monoclonal antibody or its fragment specifically bound with GDF8 and not bound with BMP11, a polynucleotide coding it, an expression vector and a host cell for antibody expression. There are studied: a method for producing the GDF8-specific antibody, a method for GDF8 presence test, a method of treating a GDF8-associated disorder.

EFFECT: use of the invention provides the new GDF8-specific antibodies that can find further application in the therapy of the GDF8-mediated diseases.

22 cl, 33 dwg, 8 tbl, 7 ex

 

The text descriptions are given in facsimile form.

1. Monoclonal antibody or its fragment, which(th) specifically binds to GDF8, but is not associated specifically with BMP11, containing(s)
defining complementary region 1 (CDR1) of the variable domain of the heavy chain (VH)selected from SEQ ID NO:19 or SEQ ID NO:25, VH CDR2 selected from SEQ ID NO:20 or SEQ ID NO:26, and VH CDR3 selected from SEQ ID NO:21 or SEQ ID NO:27; and
defining complementary region 1 (CDR1) of the variable domain of the light chain (VL)selected from SEQ ID NO:22 or SEQ ID NO:28, VL CDR2 selected from SEQ ID NO:23 or SEQ ID NO:29 and a VL CDR3 selected from SEQ ID NO:24 or SEQ ID NO:30.

2. Monoclonal antibody or its fragment according to claim 1, containing(d) a heavy chain comprising the amino acid sequence selected from the group SOS is oasa of the amino acid sequence of SEQ ID NO:14 and the amino acid sequence of SEQ ID NO:17.

3. Monoclonal antibody or its fragment according to claim 1, containing(d) a light chain comprising the amino acid sequence selected from the group consisting of the amino acid sequence of SEQ ID NO:16 and amino acid sequence of SEQ ID NO:18.

4. Monoclonal antibody or its fragment according to claim 1, containing(d) a light chain comprising the amino acid sequence of SEQ ID NO:16 and a heavy chain comprising the amino acid sequence of SEQ ID NO:14.

5. Monoclonal antibody or its fragment according to claim 1, containing(d) a light chain comprising the amino acid sequence of SEQ ID NO:18 and a heavy chain comprising the amino acid sequence of SEQ ID NO:17.

6. Monoclonal antibody or fragment according to any one of claim 2, 4 or 5 where the specified heavy chain also contains the constant domain of the heavy chain selected from the group consisting of the constant domains of the heavy chain of IgA1, IgA2, IgE, IgD, IgG1, IgG2, IgG3, IgG4 and IgM.

7. Monoclonal antibody or fragment according to any one of p, 4 or 5 where the specified light chain also contains the constant domain of the light chain selected from the group consisting of the constant domains of the light chain lambda and Kappa.

8. Monoclonal antibody or fragment according to any one of claims 1 to 7, which(th) is a fully humanized.

9. Selected polynucleotide that encodes the antibody or fragment according to any one of claims 1 to 8.

10. Selected polynucleotide according to claim 9, containing nukleinovokisly sequence of SEQ ID NO:13 encoding a variable domain of the heavy chain (VH) RK22.

11. Selected polynucleotide according to claim 9, containing nukleinovokisly sequence of SEQ ID NO:15 encoding a variable domain of the light chain (VL) RK22.

12. The expression vector containing the regulatory sequence functionally related polynucleotide according to claim 9.

13. A host cell of a mammal for expression GDF8-specific monoclonal antibody or its fragment containing the vector according to item 12.

14. A method of obtaining a GDF8-specific monoclonal antibody or its fragment comprising culturing the host cell according to item 13 and the allocation of GDF8-specific monoclonal antibody or its fragment expressed by the specified cell of the host.

15. Selected GDF8-specific monoclonal antibody or its fragment obtained(th) the method according to 14.

16. The method of determining the presence of GDF8 in the sample from the subject, comprising the stage of:
(a) bringing into contact of the sample with GDF8-specific antibody for capture;
(b) bringing into contact of the sample with GDF8-specific antibody for detection; and
(b) detecting specific binding of the antibodies for detection with GDF8, specifically associated with the antibody for capture,
where either the antibody for the capture of the, either the antibody for detection, but not both, is an antibody according to claim 1; and
where detection of specific binding indicates the presence of GDF8 in the sample.

17. The method according to clause 16, where an alternative antibody that is not an antibody according to claim 1, is RK35 antibody or antigen-binding fragment.

18. The method according to item 16 or 17, where the stage of (a) bringing into contact of the sample with GDF-specific antibody for capture is carried out in acidic conditions.

19. The method according to p, where stage (a) is carried out at a pH below about 3,0.

20. The method according to item 16 or 17, which further includes a step quantitative determination of the level of specific binding of the antibodies for detection with GDF8, whereby to determine the number of GDF8 in the specified pattern.

21. The method of treatment GDF8-associated disorder in a mammal, comprising an introduction to the specified mammal amount of GDF8-specific monoclonal antibody or its fragment according to any one of claims 1 to 8, is effective for the treatment of this disorder.

22. The use of pharmaceutical compositions in the manufacture of drugs for the treatment of GDF8-associated disorder in a mammal, where the specified pharmaceutical composition comprises a pharmaceutically acceptable carrier and GDF8-specific monoclonal antibody or fragment according to any one of claims 1 to 8.



 

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