Method of concentrating antibody preparations

FIELD: chemistry; biochemistry.

SUBSTANCE: invention pertains to bioengineering. The method involves successive steps for ultrafiltration of a first antibody preparation to obtain a second antibody preparation, diafiltration of the second antibody preparation to obtain an intermediate preparation and second ultrafiltration of the intermediate preparation to obtain a third antibody preparation. All steps are carried out at temperature ranging from approximately 30°C to approximately 50°C.

EFFECT: design of an efficient method of concentrating antibody preparations.

52 cl, 25 dwg, 25 tbl, 12 ex

 

The text descriptions are given in facsimile form.

1. Method of preparation of highly concentrated composition antibodies containing:
(a) a first ultrafiltration first drug antibodies to receive a second drug antibodies;
(b) diafiltration second drug antibodies for intermediate diafiltration drug antibodies; and
(c) a second ultrafiltration intermediate drug antibodies to receive a third drug antibodies in which the above-mentioned stage of the process performed at a temperature from about 30°to about 50°C.

2. The method according to claim 1, wherein the antibody is an anti-IgE antibody.

3. The method according to claim 1, in which one or several stages, the first ultrafiltration, the second ultrafiltration and diafiltration performed at a temperature of from about 35°to about 50°C.

4. The method according to claim 1, in which one or several stages, the first ultrafiltration, the second ultrafiltration and diafiltration performed at a temperature of from about 45±5°C.

5. The method according to claim 1, in which one or several stages, the first ultrafiltration, the second ultrafiltration and diafiltration performed at a temperature of about 45°C.

6. The method according to claim 1, in which stage (a), (b) and (C) you shall anaut at a temperature of from about 40°C to about 50°C.

7. The method according to claim 1, wherein the first drug antibody has a concentration of from about 0.1 to about 10 g/L.

8. The method according to claim 1, wherein the first drug antibody has a concentration of from about 1 g/l to about 5 g/L.

9. The method according to claim 1, wherein the second drug antibody has a concentration from about 10 g/l to about 50 g/l

10. The method according to claim 1, wherein the second drug antibody has a concentration of from about 20 g/l to about 50 g/l

11. The method according to claim 1, wherein the second drug antibody has a concentration of from about 20 g/l to about 40 g/L.

12. The method according to claim 1, wherein the third drug antibody has a concentration of from about 50 g/l to about 250 g/l

13. The method according to claim 1, wherein the third drug antibody has a concentration of from about 100 g/l to about 230 g/l

14. The method according to claim 1, wherein the third drug antibody has a concentration of from about 170 g/l to about 200 g/L.

15. The method according to claim 1, in which the intermediate drug antibody and a third drug antibodies contain retentate ultrafiltration.

16. The method according to claim 1, in which the intermediate drug antibody has a concentration of from about 25 g/l to about 35 g/l, and a third drug antibody has a concentration of from about 170 g/l to about 200 g/L.

17. The method according to claim 1, in which the first ultrafiltration first drug antibodies provides a second drug EN is the body, having a concentration of about 30 g/l, and the second ultrafiltration concentrates intermediate drug antibodies to provide a third drug antibody having a concentration of from about 170 to about 200 g/L.

18. The method according to claim 1, in which the process is performed for from about 1 hour to about 10 hours

19. The method according to claim 1, in which the process is from about 2 hours to about 5 hours

20. The method according to claim 1, in which the process is performed for approximately 3 hours

21. The method according to claim 1, in which the first and second ultrafiltration is carried out using the ultrafiltration membrane having a nominal pore size corresponding to a molecular weight from about 5 kDa to about 50 kDa.

22. The method according to claim 1, in which the first and second ultrafiltration is carried out using the ultrafiltration membrane having a nominal pore size corresponding to a molecular weight of from about 10 kDa to about 30 kDa.

23. The method according to claim 1, wherein the first drug antibodies includes an antibody having an average molecular weight from about 100 kDa to about 200 kDa.

24. The method according to claim 1, wherein the first drug antibodies includes an antibody having an average molecular weight of approximately 150 kDa.

25. The method according to claim 1, wherein stages (a), (b) and (C) using membrane ultrafiltration.

26. The method according to claim 1, wherein the first ultrafilter the Oia and the second ultrafiltration is performed with the same membrane ultrafiltration.

27. The method according to claim 1, wherein the ultrafiltration membrane used on stage filtration (a), is used in stage (b) and stage (C).

28. The method according to claim 1, wherein the first ultrafiltration, the second ultrafiltration and diafiltration carried out in the mode of tangential flow towards the membrane ultrafiltration.

29. The method according to claim 1, wherein the first ultrafiltration, the second ultrafiltration and diafiltration carried out in the mode of tangential flow in relation to the same membrane ultrafiltration.

30. The method according to claim 1, wherein the first ultrafiltration, the second ultrafiltration is carried out by membrane ultrafiltration with composite regenerated cellulose.

31. The method according to claim 1, in which diafiltration do with the replacement of the buffer at constant volume or at constant concentration, or may use a combination of both modes.

32. The method according to claim 1, in which the replacement of the buffer solution when diafiltration carried out with a ratio of from about 5 to about 15.

33. The method according to claim 1, in which the replacement volume of buffer solution at diafiltration carry about 8 times.

34. The method according to claim 1, wherein stage (b) comprises more than one stage of diafiltration.

35. The method according to clause 34, in which the first stage of diafiltration includes replacement of buffer solution at diafiltration primerno times, and the second stage of diafiltration includes replacement of buffer solution at diafiltration about 4 times.

36. The method according to claim 1, wherein when diafiltration first buffer solution replace the second buffer solution.

37. The method according to claim 1, wherein when diafiltration use buffer solution having a composition that differs from the composition of the buffer solution in stage (a).

38. The method according to p, in which the first buffer solution contains a mixture of aqueous sodium chloride and Tris-buffer.

39. The method according to p, in which the second buffer solution includes a mixture of an aqueous solution of histidine chloride and arginine chloride.

40. The method according to claim 1, in which the output of the third drug antibodies greater than about 70 wt.% the weight of the antibody in the first drug antibodies.

41. The method according to claim 1, in which the output of the third drug antibody is from about 80 to 100 wt.% the weight of the antibody in the first drug antibodies.

42. The method according to claim 2, in which the output of the third drug antibodies greater than about 98 wt.% the weight of the antibody in the first drug antibodies.

43. The method according to claim 1, wherein the first ultrafiltration is carried out at the intensity of recirculation from approximately 0.5 l/min/ft2to about 5 l/min/ft2.

44. The method according to claim 1, wherein the ultrafiltration and diafiltration carried out at a transmembrane pressure of from about 10 psi to the ome 50 psi.

45. The method according to claim 1, wherein the ultrafiltration and diafiltration carried out at a transmembrane pressure of from about 10 psi to about 50 psi.

46. The method according to claim 1, in which the bandwidth for stage (C) is about 80 g/ft2/h to about 120 g/ft2/h, about 135 g/ft2/h, approximately 145 g/ft2/h, about 175 g/ft2/h to about 180 g/ft2/h, about 265 g/ft2/h, about 285 g/ft2/h or approximately 290 g/ft2/PM

47. The method according to claim 1, in which the aggregated level of pollutants is less than 2 wt.%.

48. The method according to claim 1, in which the concentration of the antibody with a detectable amount of bio-contaminants less than about 100 CFU/ml

49. The method according to claim 1, wherein the third drug antibodies is the number of detectable bio-contaminants of about 18, CFU/ml.

50. The method according to claim 1, wherein the third drug antibodies is the number of detectable bio-contaminants order of 0.13 CFU/ml

51. The method according to claim 1, wherein the first drug antibodies purified prior to stage (a).

52. The method according to claim 2, in which the antibody contained in the first drug antibodies, has a purity of about 99.8 per cent prior to stage (a).



 

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