The antibody against the protein, parathyroid related hormone human dna, vector, method of production and use of antibodies

 

(57) Abstract:

The invention relates to biotechnology, in particular to antibodies against Rtln, obtained by genetic engineering methods. The invention relates to the creation of chimeric L - and H-chains and full of antibodies based on them, humanized antibodies and polypeptides of the V region of the L - and H-chains; DNA encoding them, and vector. Further, the invention discloses methods of obtaining chimeric and gumanitarnogo antibodies and use in pharmaceutical compositions for the prevention and treatment of weight loss, associated in particular with malignancy. An antibody-based tool created to suppress hypercalcemia, to improve the condition when hypophosphatemia. The invention allows to obtain humanized antibodies, characterized by sufficient activity of binding and neutralizing specific antigen (RTRN), which is ensured through the creation of effective methods of obtaining them. 26 C. and 25 C.p. f-crystals, 31 tab., 6 table.

Description text in facsimile form. T T T,

1. Chimeric L-chain comprising a C-area L-chain of human antibodies and V-region L-chain mouse monoclonal antibodies against the protein, parathyroid related hormone >/P>2. Chimeric L-chain under item 1, characterized in that the region is a P-chain.

3. Chimeric H-chain comprising P-region N-chains of human antibodies and the V region of H chain mouse monoclonal antibodies against the protein, parathyroid related hormone human, in which the V region of H chain comprises the amino acid sequence shown in SEQ ID NO:46.

4. Chimeric H-chain according to p. 3, characterized in that the region is C1-circuit.

5. Chimeric monoclonal antibody against the protein, parathyroid related hormone human, comprising the chimeric L-chain under item 1 or 2 and chimeric H-chain under item 1 or 2 and chimeric H-chain under item 3 or 4.

6. Polypeptide comprising V region of the L-chain gumanitarnogo antibodies against the protein, parathyroid related hormone human frame region 1-3 specified V-region L-chain derived from the V region of the L-chain of human antibodies HSU03868, and a frame region 4 of the same V region of the L-chain is obtained from the V region of the L-chain of human antibodies S25755, and hypervariable sections 1-3 of the specified V-region L-chain comprise the amino acid sequence selected from the group of sequences SEQ ID nos:59-61.

7. Polypeptide comprising V region of the L-chain humanize the marks V-region L-circuit is essentially identical to frame areas 1-3 human antibodies HSU03868, and a frame region 4 of the same V region of the L-chain is essentially identical to the framework region 4 human antibodies S25755, and hypervariable sections 1-3 of the specified V-region L-chain comprise the amino acid sequence selected from the group of sequences SEQ ID nos:59-61.

8. The polypeptide under item 7, characterized in that the 36th and 49th amino acids of frame areas on the definition of Kabat are, respectively, tyrosine and aspartic acid.

9. The polypeptide under item 8, characterized in that it includes any one amino acid sequence shown in SEQ ID nos:48-51.

10. The polypeptide under item 7, characterized in that the 87th amino frame areas on the definition of Kabat is an isoleucine.

11. The polypeptide under item 7, characterized in that 45-I and 87th amino acids of frame areas on the definition of Kabat are, respectively, lysine and isoleucine.

12. The polypeptide under item 10, characterized in that it includes any one amino acid sequence shown in SEQ ID nos:52-55.

13. The polypeptide according to p. 11, characterized in that it comprises the amino acid sequence shown in SEQ ID NO:52 or 54.

14. Polypeptide comprising V region of H chain is STI 1-4 specified V-N-chain derived from V-N-chain of human antibodies human subgroup III, and hypervariable sections 1-3 of the specified V-N-chain comprise the amino acid sequence selected from the group of sequences SEQ ID nos:62-64.

15. Polypeptide comprising V region of H chain gumanitarnogo antibodies against the protein, parathyroid related hormone human frame region 1-4 specified V-N-chain is essentially identical to frame areas 1-4 human antibodies human subgroup III, and hypervariable sections 1-3 of the specified V-N-chain comprise the amino acid sequence selected from the group of sequences SEQ ID nos:62-64.

16. The polypeptide under item 14 or 15, wherein the human antibody is an antibody of human S31679.

17. The polypeptide under item 14, characterized in that it comprises the amino acid sequence shown in SEQ ID NO:56.

18. L-chain gumanitarnogo antibodies against the protein, parathyroid related hormone human, comprising a polypeptide comprising the C-region L-chain of human antibodies and the polypeptide according to any one of paragraphs.6-13.

19. L-chain gumanitarnogo antibodies under item 18, characterized in that the region is a P-chain.

20. L-chain gumanitarnogo antibodies frame areas 1-3 human antibodies HSU03868, accordingly, the frame region 4 is essentially identical to the framework region 4 human antibodies S25755, and hypervariable sections 1-3 include amino acid sequences shown, respectively, in SEQ ID nos:59-61.

21. H-chain gumanitarnogo antibodies against the protein, parathyroid related hormone human, comprising a polypeptide comprising a P-region N-chains of human antibodies and the polypeptide according to any one of paragraphs.14-17.

22. H-chain gumanitarnogo antibodies for p. 21, characterized in that the region is C1-circuit specified frame region 1-4 allocated, respectively, of the frame regions 1-4 human antibodies human subgroup III, and hypervariable sections 1-3 include amino acid sequences shown, respectively, in SEQ ID nos:62-64.

23. Humanitariannet antibody against the protein, parathyroid related hormone human, comprising the L-chain gumanitarnogo antibody according to any one of paragraphs.18-20 and H-chain gumanitarnogo antibodies on PP.21 or 22.

24. DNA comprising the sequence of bases encoding a V region of the L-chain mouse monoclonal antibodies against the protein, parathyroid related hormone human, in which V is ª the sequence of bases encoding V region of the L-chain mouse monoclonal antibodies against the protein, parathyroid related hormone human, in which the sequence of bases encoding a V region of the L-chain shown in SEQ ID NO:65.

26. DNA comprising the sequence of bases encoding a V region of H chain mouse monoclonal antibodies against the protein, parathyroid related hormone human, in which the V region of H chain comprises the amino acid sequence shown in SEQ ID NO:46.

27. DNA comprising the sequence of bases encoding a V region of H chain mouse monoclonal antibodies against the protein, parathyroid related hormone human, in which the sequence of bases encoding a V region of H chain shown in SEQ ID NO:57.

28. DNA encoding a chimeric L-chain under item 1 or 2.

29. DNA under item 28, wherein the DNA encoding a chimeric L-chain comprises the sequence of bases shown in SEQ ID NO:65.

30. DNA encoding a chimeric H-chain under item 3 or 4.

31. DNA under item 30, wherein the DNA encoding a chimeric H chain comprises the sequence of bases shown in SEQ ID NO:57.

32. DNA comprising the base sequence is Sevani, shown in SEQ ID NO:66-74.

34. DNA comprising the sequence of bases encoding the polypeptide according to any one of paragraphs.14-17.

35. DNA in p. 34, comprising a sequence of bases shown in SEQ ID NO:58.

36. DNA encoding the L-chain gumanitarnogo antibody according to any one of paragraphs. 18-20.

37. DNA encoding the L-chain gumanitarnogo antibody comprising the sequence of bases encoding any one of the amino acid sequence shown in SEQ ID NO:47-55.

38. DNA under item 37, wherein the DNA of the L-chain gumanitarnogo antibody comprises any one of the sequence of bases shown in SEQ ID NO:66-74.

39. DNA encoding the H-chain gumanitarnogo antibodies under item 21 or 22.

40. DNA N-chain gumanitarnogo antibody comprising the sequence of bases encoding amino acid sequence shown in SEQ ID NO:56.

41. DNA under item 40, wherein the DNA N-chain gumanitarnogo antibody comprises the sequence of bases shown in SEQ ID NO:58.

42. Recombinant vector comprising the DNA according to any one of paragraphs.24-41.

43. Recombinant vector according to p. 42, characterized in that it is embedded in the transform is kiuchumi cultivation transformant, transformed expressing a vector comprising the DNA according to any one of paragraphs.24, 25, 28 and 29 and expressing a vector comprising the DNA according to any one of paragraphs.26, 27, 30 and 31, and the allocation of the resulting culture chimeric antibodies against the protein, parathyroid related hormone human.

45. The method of obtaining gumanitarnogo antibodies against the protein, parathyroid related hormone human, including the cultivation of transformant transformed by expressing a vector comprising the DNA according to any one of paragraphs.32, 33, 36-38, and expressing a vector comprising the DNA according to any one of paragraphs. 34, 35, 39-41, and the allocation of the resulting culture gumanitarnogo antibodies against the protein, parathyroid related hormone human.

46. Pharmaceutical composition for prevention or treatment of weight loss for the prevention or improvement of reduced food intake, anorexia, nausea, metabolic alkalosis, broken blood pH, for regulating the temperature changes of the body, for the prevention or improvement of decreased body temperature, decreased spontaneous activity, reduced activity, reduced QOL of the patient in connection with a malignant tumour, and to suppress hypercalcemia the th media.

47. Means for suppression of hypercalcemia or to improve upon hypophosphatemia, including humanitariannet antibody on p. 23 as an active ingredient.

48. Means on p. 47, characterized in that the hypercalcemia due to malignancy.

49. Means on p. 48, wherein the malignant tumor is at least one selected from the group including pancreatic cancer, lung cancer, pharynx cancer, larynx cancer, tongue cancer, cancer of the gums, esophagus cancer, stomach cancer, cancer of the bile ducts, breast cancer, kidney cancer, bladder cancer, uterine cancer, prostate cancer and malignant lymphoma.

50. Means on p. 47, characterized in that hypophosphatemia is hypophosphatemic rickets.

51. Means on p. 47, characterized in that hypophosphatemia is hypophosphatemic rickets resistant to vitamin D.

 

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