Humanized antibodies that recognize the verotoxin ii, and producing their line of cells

 

The invention relates to medicine and relates to humanized antibodies that recognize the verotoxin II, and producing their line of cells. The invention consists in creating gumanitarnogo antibodies, representing humanitarian form antibodies VTm. 1 mouse, which is characterized by a variable region light chain and the variable region of the heavy chain and which specifically binds and neutralizes VT2 or its variant, and also relates to pharmaceutical compositions and methods of treatment using the indicated antibodies. The advantage of the invention is to obtain antibodies that are not immunogenic to humans, but with the properties of the antibodies of the mouse, which can be obtained by methods suitable for therapeutic use. 5 C. and 20 C. p. F.-ly, 6 PL. , 7 Il.

Tabletif

Claims

1. Humanitariannet antibody representing humanitarian form antibodies VTm1-1 mouse, with the indicated antibody mouse is characterized by a variable region of the light chain shown in Fig.1B, and the variable region of the heavy chain shown in Fig.1A, where the specified humanitariannet antibody with the demonstrates with antibody VTm1-1 mice for specific binding to VT2 and/or VT2 variant.

3. Humanitariannet antibody under item 1, which specifically binds to a subunit In VT2 and/or subunit In VT2 variant.

4. Humanitariannet antibody under item 1 or 2, including region complementarity determining, from antibodies VTm1-1 mouse, and the frames of variable regions of the heavy and light chains from the frames of the heavy and light chains of antibodies GF4 person, provided that at least one position selected from the group consisting of L49, N, N, N and N, are occupied by the amino acid present at the corresponding position of the frame variable regions of the heavy or light chain antibodies VTm1-1 mouse moreover, the specified humanitariannet antibody specifically binds the verotoxin II with an affinity constant between 107M-1and ten times greater affinity than such antibodies VTm1-1 mouse.

5. Humanitariannet antibody under item 4, where each position selected from the group consisting of L49, S, H30, N and N, are occupied by the amino acid present at the corresponding position of the frame variable regions of the heavy or light chain antibodies VTm1-1 mouse.

6. Humanitariannet antibody under item 5, where at least one position selected from the group L3, L4, L19, L76, L79, L85, H1, H4, H5, N, N and N, are occupied by the amino acid prisutstvuushih>7. Humanitariannet antibody under item 6, where each position selected from the group L3, L4, L19, L76, L79, L85, H1, H4, H5, N, N and N, are occupied by the amino acid present at the corresponding position of the consensus sequence of the heavy or light chains of human antibodies.

8. Humanitariannet antibody under item 1 or 2, containing the variable region of the heavy chain shown in Fig.2A, and the variable region of the light chain shown in Fig.2B, provided that one or more positions selected from the group consisting of L49, N, N, N, N, L3, L4, L19, L76, L79, L85, H1, H4, H5, N, N and N, can be replaced, as shown in tables 2 and 3.

9. Humanitariannet antibody under item 1 or 2, containing the variable region of the heavy chain shown in Fig.2A, and the variable region of the light chain shown in Fig.2B.

10. Humanitariannet antibody under item 1 or 2, containing humanitarian heavy chain having a sequence at least 85% identical humanized heavy chain shown in Fig.2A, and humanitarian light chain having a sequence at least 85% identical to the humanized light chain shown in Fig.2B, provided that at least one position selected and frame variable regions of the heavy or light chain antibody mouse VTm1-1.

11. Humanitariannet antibody under item 1 or 2, where the antibody contains two pairs of dimers of light/heavy chains, where each chain contains a variable region and a constant region.

12. Humanitariannet antibody under item 1 or 2, which is a Fab fragment or F(ab')2.

13. Humanitariannet antibody under item 1 or 2 in purified form.

14. Humanitariannet antibody under item 1 or 2, which has the immunoglobulin isotype IgG1.

15. The method of obtaining gumanitarnogo antibodies VTm1-1, providing for the cultivation of cell lines, which encodes the heavy and light chain gumanitarnogo antibody described in any of paragraphs.1-14, the result is expressed humanitariannet antibody, and the allocation of gumanitarnogo antibodies expressed by the specified cell line.

16. The method according to p. 15, further providing for mixing the antibody with a pharmaceutically acceptable carrier to obtain a pharmaceutical composition.

17. Pharmaceutical composition for the treatment of infections caused by E. Li, verotoxin producing, and haemolytic uraemic syndrome containing humanitariannet antibody described in any of paragraphs.1-14, and a pharmaceutically premliminary region of the heavy chain, presented on Fig.2A, and the variable region of the light chain shown in Fig.2B.

19. A method of treating a patient at risk or suffering from the toxic effects of verotoxin, including the introduction of patient effective dose gumanitarnogo antibodies, representing humanitarian form antibodies VTm1-1 mouse, with the indicated antibody mouse is characterized by a variable region of the light chain shown in Fig.1B, and the variable region of the heavy chain shown in Fig.1A, where the specified humanitariannet antibody specifically binds and neutralizes subunit In VT2 and/or subunit In VT2 variant.

20. The method according to p. 19, in which the antibody competes with the antibody VTm1-1 mice for specific binding of verotoxin II or option verotoxin II.

21. The method according to p. 19, in which the antibody is humanitariannet antibody containing the variable region of the heavy chain shown in Fig.2A, and the variable region of the light chain shown in Fig.2B.

22. The method according to p. 19, wherein said patient is infected with E. coli, verotoxin producing, and the antibody is administered therapeutically.

23. The method according to p. 22, additionally providing for the monitoring with the.

24. The method according to p. 19, wherein said patient is susceptible E. coli, verotoxin producing, and the antibody is administered prophylactically.

25. Cell line murine myeloma producing the antibody described in any of paragraphs.1-14.

 

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