Process for isolation of plasma or serum proteins

1. A process for the large-scale isolation of one or more proteins from a protein solution, wherein the protein solution is obtained from a source selected from the group consisting of human blood and a solution derived from human blood, and wherein the protein solution has not been supplemented with an alcohol, the process comprising the steps of:

a) adjusting the pH of the solution to a pH in the range of 3 to 10;

b) adjusting the ionic strength or conductivity of the protein solution to an ionic strength in the range of 0.0001 to 12 or a conductivity in the range of 0.01 to 1000 mS/cm;

c) applying said protein solution to an adsorption column comprising an adsorbent, said adsorbent comprises a particle with at least one high density non-porous core, surrounded by a porous material, the adsorbent comprises a particle density of at least 1.5 g/ml and a mean volume particle diameter of at most 150 μm, whereby one or more proteins are bound to the adsorbent;

d) obtaining a non-bound material fraction from the column, which non-bound material fraction contains at least one non-bound protein;

e) subjecting the adsorbent to at least one elution buffer to elute at least one of the proteins bound to the adsorbent, thereby obtaining a bound material fraction; and

f) subjecting the non-bound material fraction to further processing to obtain at least one non-bound protein therefrom.

2. The process according to claim 1 , wherein the proteins are human blood proteins from the human blood solution.

3. The process according to claim 1 , further comprising at least one virus elimination treatment.

4. The process according to claim 3 , wherein the at least one virus elimination treatment is performed prior to contacting the protein solution with the adsorbent.

5. The process according to claim 3 , wherein the virus elimination treatment involves addition of detergent and/or an organic solvent to the protein solution.

6. The process according to claim 1 , wherein the adsorbent comprises a functionalized matrix polymer carrying a plurality of covalently attached functional groups comprising an aromatic ring-system, a heteroaromatic ring-system or one or more acidic groups.

7. The process according to claim 2 , wherein the human blood proteins are selected from the group consisting of albumin, IgG, IgA, IgM, IgD, IgE, alpha-1-proteinase inhibitor, blood pro-coagulation protein, blood anti-coagulation protein, anti-angiogenic protein, α-2-antiplasmin, C-1 esterase inhibitor, apolipoprotein, HDL, LDL, Fibronectin, beta-2-glycoprotein I, fibrinogen, plasminogen, plasmin, plasminogen activator, plasminogen inhibitor, plasma protease inhibitor, anti-thrombin III, streptokinase, inter-alpha-trypsin inhibitor, α-2-macroglobulin, amyloid protein, ferritin, pre-albumin, GC-globulin, haemopexin, C3-complement , transferrin, urokinase and α-1-acid-glycoprotein.

8. The process according to claim 1 , wherein, after applying the protein solution to the adsorption column, one or more proteins are washed out as a non-bound protein with one or more washing buffers.

9. The process according to claim 1 , wherein one non-bound protein is an alpha-1-proteinase inhibitor.

10. The process according to claim 1 , wherein, prior to applying the protein solution to the adsorption column, the pH of the protein solution is adjusted to a pH in the range of pH 3.0 to pH 10.0.

11. The process according to claim 10 , wherein the pH-value for capturing and for elution is maintained in the range of pH 3.0 to pH 10.0.

12. The process according to claim 1 , wherein, prior to applying the protein solution to the adsorption column, the ionic strength is adjusted.

13. The process according to claim 1 , wherein, prior to applying the protein solution to the adsorption column, the conductivity is adjusted.

14. The process according to claim 1 , wherein the flow rate of the protein solution through the column is at least 2 cm/min.

15. The process according to claim 1 , wherein the adsorbent has a dynamic binding capacity at 10% break-through for said at least one protein of at least 5 g per liter sedimented adsorbent.

16. The process according to claim 1 , wherein at least 2 proteins are isolated from the protein solution, wherein the proteins are selected from the group consisting of albumin, IgG, IgA, IgM, IgD, IgE, alpha-1-proteinase inhibitor, blood pro-coagulation protein, blood anti-coagulation protein, anti-angiogenic protein, α-2-antiplasmin, C-1 esterase inhibitor, apolipoprotein, HDL, LDL, Fibronectin, beta-2-glycoprotein I, fibrinogen, plasminogen, plasmin, plasminogen activator, plasminogen inhibitor, plasma protease inhibitor, anti-thrombin III, streptokinase, inter-alpha-trypsin inhibitor, α-2-macroglobulin, amyloid protein, ferritin, pre-albumin, GC-globulin, haemopexin, C3-complement, transferrin, urokinase and α-1-acid-glycoprotein, and wherein the proteins are isolated in at least 2 fractions.

17. The process according to claim 16 , wherein at least 2proteins are isolated from the protein solution, wherein the proteins are selected from the group consisting of α-1 proteinase inhibitor, IgG, human albumin, transferrin, thrombin, Factor II, Factor V, Factor VII, Factor VII, Factor IX, protein C, Protein S, α-1-acid-glycoprotein and fibrinogen, and wherein the proteins are isolated in at least 2fractions.

18. The process according to claim 1 , wherein at least one of the proteins bound to the adsorbent comprises fibrinogen and wherein the at least one non-bound protein comprises Factor II, Factor V, Factor VII, Factor VIII, von Willebrand factor, Factor VIII - von Willebrand factor complex, Factor IX, Factor X, Factor XI, C1 inhibitor, protein C and/or Protein S.

19. The process according to claim 1 , wherein the bound proteins are albumin and IgG and the non-bound protein is α-1-proteinase inhibitor.

20. The process according to claim 19 , wherein albumin and IgG are isolated from the bound fraction by stepwise elution.

21. The process according to claim 1 , wherein fibrinogen and IgG are bound to the adsorbent and simultaneously albumin may be obtained as non-bound material from the adsorbent.

22. The process according to claim 21 , wherein fibrinogen and IgG may be obtained from the adsorbent by stepwise elution.

23. The process according to claim 1 , wherein the bound proteins are fibrinogen, albumin and IgG and the non-bound protein is α-1-proteinase inhibitor.

24. Theprocess according to claim 23 , wherein fibrinogen, albumin and IgG are obtained from the adsorbent by stepwise elution.

25. The process according to claim 1 , wherein at least 2 of fibrinogen, albumin, and/or IgG are bound to the adsorbent and simultaneously α-1-acid glycoprotein is obtained as non-bound material from the adsorbent.

26. The process according to claim 25 , wherein the at least 2of fibrinogen, albumin and/or IgG are obtained from the adsorbent by stepwise elution.

27. The process according to claim 1 , wherein the bound proteins are at least 2 of fibrinogen, albumin and/or IgG and the non-bound proteins are α-1-acid glycoprotein and/or α-1-proteinase inhibitor.

28. The process according to claim 27 , wherein the at least 2 of fibrinogen, albumin and/or IgG may be obtained from the adsorbent by stepwise elution.

29. A process for the large-scale isolation of one or more proteins from a protein solution, wherein the protein solution is obtained from a source selected from the group consisting of human blood and a solution derived from human blood, and wherein the protein solution has not been supplemented with an alcohol, said process comprising the steps of:

a) adjusting the pH of the protein solution to a pH in the range of 3 to 10;

b) adjusting the ionic strength or conductivity of the protein solution to an ionic strength in the range of 0.0001 to 12 or a conductivity in the range of 0.01 to 1000 mS/cm;

c) applying the protein solution to an adsorption column comprising an adsorbent, wherein the adsorbent comprises a functionalised matrix polymer carrying a plurality of covalently attached functional groups comprising an aromatic or heteroaromatic ring-system and one or more acidic groups, whereby one or more proteins are bound to the adsorbent;

d) obtaining a non-bound material fraction from the column, which non-bound material fraction contains at least one non-bound protein;

e) subjecting the adsorbent to at least one elution buffer to elute at least one of the proteins bound to the adsorbent; and

f) subjecting the non-bound material fraction to further processing to isolate at least one non-bound protein therefrom.

30. The process according to claim 29 , further comprising at least one virus elimination treatment.

31. The process according to claim 29 , wherein one of the non-bound proteins is α-1-proteinase inhibitor.

32. The process according to claim 29 , wherein one of the bound proteins is human albumin.

33. The process according to claim 29 , wherein one of the bound proteins is fibrinogen.

34. The process according to claim 29 , wherein one of the bound proteins is transferrin.

35. The process according to claim 29 , wherein one of the non-bound proteins is α-1-acid glycoprotein.

36. The process according to claim 29 , wherein one of the non-bound proteins is a coagulation or anticoagulation factor.

37. The process according to claim 36 , wherein the coagulation or anti-coagulation factor is selected from the group consisting of Factor II, Factor V, Factor VII, Factor VIII, von Willebrand factor, Factor VIII - von Willebrand factor complex, Factor IX, Factor X, Factor XI, C1 inhibitor, protein C and Protein S.

38. A process for the large-scale isolation of one or more proteins from a protein solution, wherein the protein solution is obtained from a source selected from the group consisting of human blood and a solution derived from human blood, and wherein the protein solution has not been supplemented with an alcohol, said process comprising the steps of:

a) providing a protein solution comprising one or more proteins obtained from a source selected from the group consisting of human blood or a solution derived from human blood, the protein solution having a preset pH and a preset ionic strength or conductivity;

b) contacting the protein solution with an adsorbent, wherein the adsorbent comprises a particle with at least one high density non-porous core, surrounded by a porous material, whereby one or more proteins is bound to the adsorbent;

c) subjecting the adsorbent to an elution buffer to elute at least one of the proteins bound to the adsorbent; and

d) subjecting the adsorbent to one or more additional elution buffers to elute one or more remaining proteins bound to the adsorbent.

39. The process according to claim 38 , further comprising at least one virus elimination treatment.

40. The process according to claim 38 , wherein one of the proteins isolated from the protein solution is an α-1-proteinase inhibitor.

41. The process according to claim 38 , wherein one of the proteins isolated from the protein solution is human albumin.

42. The process according to claim 38 , wherein one of the proteins isolated from the protein solution is fibrinogen.

43. The process according to claim 38 , wherein one of the proteins isolated from the protein solution is transferrin.

44. The process according to claim 38 , wherein one of the proteins isolated from the protein solution is α-1- acid-glycoprotein.

45. The process according to claim 38 , wherein one of the proteins isolated from the protein solution is a coagulation or anticoagulation factor.

46. The process according to claim 45 , wherein the coagulation or anti-coagulation factor is selected from the group consisting of Factor II, Factor V, Factor VII, Factor VIII, von Willebrand factor, Factor VIII - von Willebrand factor complex, Factor IX, Factor X, Factor XI, C1 inhibitor, protein C and Protein S.

47. The process according to claim 1 , wherein the protein solution is serum and/or plasma.

48. The process according to claim 2 , wherein the human blood proteins are human plasma proteins or human serum proteins.

49. The process according to claim 16 , wherein 3 proteins are isolated from the protein solution, and wherein the 3 proteins are isolated in 3 fractions.

50. The process according to claim 16 , wherein 4 proteins are isolated from the protein solution, and wherein the 4 proteins are isolated in 4 fractions.

51. The process according to claim 16 , wherein 5 proteins are isolated from the protein solution, and wherein the 5 proteins are isolated in 5 fractions.

52. The process according to claim 16 , wherein 6 proteins are isolated from the protein solution, and wherein the 6 proteins are isolated in 6 fractions.

53. The process according to claim 17 , wherein 3 proteins are isolated from the protein solution, and wherein the 3 proteins are isolated in 3 fractions.

54. The process according to claim 17 , wherein 4 proteins are isolated from the protein solution, and wherein the 4 proteins are isolated in 4 fractions.

55. The process according to claim 17 , wherein 5 proteins are isolated from the protein solution, and wherein the 5 proteins are isolated in 5 fractions.

56. The process according to claim 17 , wherein 6 proteins are isolated from the protein solution, and wherein the 6 proteins are isolated in 6 fractions.

57. The process according to claim 25 , wherein fibrinogen, albumin and IgG are bound to the adsorbent.

58. The process according to claim 27 , wherein fibrinogen, albumin and IgG are bound to the adsorbent.

59. The process according to claim 5 , wherein the detergent and/or organic solvent is selected from the group consisting of polysorbate detergents, octylphenol ethoxylate detergents, and tri-n-butyl phosphate.

60. The process of claim 1 , wherein the human blood comprises human plasma or serum.

61. The process of claim 7 wherein the blood pro-coagulation protein or anti-coagulation protein is selected from the group consisting of Factor II, Factor V, Factor VII, Factor VIII, von Willebrand factor, Factor VIII - von Willebrand factor complex, Factor IX, Factor X, Factor XI, C1 inhibitor, protein C and Protein S.

62. The process of claim 16 wherein the blood pro-coagulation protein or anti-coagulation protein is selected from the group consisting of Factor II, Factor V, Factor VII, Factor VIII, von Willebrand factor, Factor VIII - von Willebrand factor complex, Factor IX, Factor X, Factor XI, C1 inhibitor, protein C and Protein S.