PURIFICATION OF FVIII FROM PLASMA USING SILICON OXIDE ADSORPTION

1 . A method of separating plasma cryoprecipitate comprising a blood coagulation factor and fibrinogen into a first fraction comprising a blood coagulation factor isolate and a second fraction containing the fibrinogen, the method comprising:

(a) contacting the plasma cryoprecipitate with solid Si 02 , thereby adsorbing the fibrinogen onto the solid Si 02 ; and

(b) separating the fibrinogen adsorbed onto the solid Sith from the blood factor, thereby forming the first fraction and the second fraction.

2 . The method according to claim 1 , wherein the SiO 2 is fumed SiO 2 .

3 . The method according to claim 2 , wherein the fumed SiO 2 is hydrophilic colloidal SiO 2 .

4 . The method according to claim 1 , further comprising (c), prior to (a), suspending the cryoprecipitate in water, forming a cryoprecipitate suspension.

5 . The method according to claim 4 , wherein the water is from about 15° C. to about 37° C.

6 . The method according to claim 5 , wherein the water is from about 20° C. to about 32° C.

7 . The method according to claim 4 , wherein the cryoprecipitate and water are in a ratio of from about 1:2 to about 1:7 in the cryoprecipitate suspension.

8 . The method according to claim 7 , wherein the cryoprecipitate and water are in a ratio or from about 1:3.5 to about 1:5 in the cryoprecipitate suspension.

9 . The method according to claim 4 , wherein the cryoprecipitate suspension further comprises CaCl 2 .

10 . The method according to claim 9 , wherein the CaCl 2 is present in from about 40 μM to about 50 mM in the cryoprecipitate suspension.

11 . The method according to claim 10 , wherein the CaCl 2 is present in from about 0.045 M to about 0.055 M in the cryoprecipitate suspension.

12 . The method according to claim 4 , wherein the SiO 2 is present in the suspension in from about 5 g to about 30 g of SiO 2 per kilogram of the suspension.

13 . The method according to claim 12 , wherein the SiO 2 is present in the suspension in from about 10 g to about 20 g of SiO 2 per kilogram of the suspension.

14 . The method according to claim 4 , wherein the cryoprecipitate suspension further comprises from about 2 g to about 10 g of filter aid per kilogram of cryoprecipitate suspension.

15 . The method according to claim 14 , wherein the filter aid is present in the cryoprecipitate suspension in from about 4 g to about 8 g per kilogram of the cryoprecipitate suspension.

16 . The method according to claim 4 , wherein the cryoprecipitate suspension is mixed to homogeneity.

17 . The method according to claim 16 , further comprising (d) passing the cryoprecipitate suspension through a filtration device, thereby forming a filter cake and a filtrate.

18 . The method according to claim 17 , wherein the filtration device is a mesh screen.

19 . The method according to claim 18 , wherein the mesh screen has pores of from about 100 μm to about 400 μm in diameter.

20 . The method according to claim 17 , further comprising (e) following (d), washing the filter cake with an aqueous wash solution.

21 . The method according to claim 18 , wherein the aqueous wash solution is sodium chloride solution.

22 . The method according to claim 21 , further comprising (e) filtering the filtrate through a 0.2 μm filter, forming a first filtrate.

23 . The method according to claim 20 , further comprising (f), prior to (e), adding sodium chloride to the first filtrate.

24 . The method according to claim 23 wherein the sodium chloride is added to the first filtrate to a final concentration of from about 100 mM and about 200 mM

25 . The method according to claim 22 , wherein the sodium chloride is added to the first filtrate to a final concentration of about 150 mM.

26 . The method according to claim 22 , further comprising (g), prior to (e), adding calcium chloride to the first filtrate to a final concentration of from about 0.45 M to about 0.55 M.

27 . The method according to claim 23 , wherein the calcium chloride is added to the first filtrate to a final concentration of about 0.050 M.

28 . The method according to claim 23 , further comprising (h), prior to (e), adding calcium chloride to the first filtrate.

29 . The method according to claim 1 , further comprising (i) contacting the first filtrate of (e) with a homogeneous solvent/detergent mixture, forming a first filtrate suspension.

30 . The method according to claim 29 , wherein the homogeneous solvent/detergent mixture is octoxynol and tri(n-butyl)phosphate.

31 . The method according to claim 30 , wherein the homogeneous solvent/detergent mixture is 1.0% ±0.1% (v/v) octoxynol and 0.3% ±0.03% (v/v) tri(n-butyl)phosphate.

32 . The method according to claim 29 , further comprising (j) filtering the first filtrate suspension through an aggregation removal filter, forming a second filtrate.

33 . The method according to claim 1 , wherein the blood coagulation is Factor VIII.

34 . The method according to claim 1 , wherein centrifugation is not used in separating the fibrinogen adsorbed onto the solid SiO 2 or Al(OH) 3 from the blood factor, forming the first fraction and the second fraction.

35 . The method according to claim 1 , wherein centrifugation is used in separating the fibrinogen adsorbed onto the solid SiO 2 from the blood factor, forming the first fraction and the second fraction.

36 . A blood coagulation factor isolate prepared by the method according to claim 1 .

37 . The blood coagulation factor isolate prepared by the method according to claim 1 , wherein said blood coagulation factor isolate contains less fibrinogen than the plasma cryoprecipitate.

33 . The blood coagulation factor isolate prepared by the method according to claim 1 , wherein the blood coagulation factor is FVIII.