Removal of serine proteases by treatment with finely divided silicon dioxide
The present invention provides novel methods for reducing the serine protease and/or serine protease zymogen content of a plasma-derived protein composition. Also provided are methods for manufacturing plasma-derived protein compositions having reduced serine protease and\or serine protease zymogen content. Among yet other aspects, the present invention provides aqueous and lyophilized compositions of plasma-derived proteins having reduced serine protease and/or serine protease zymogen content. Yet other aspects include methods for treating, managing, and/or preventing a disease comprising the administration of a plasma-derived protein composition having a reduced serine protease or serine protease zymogen content.
1. A method for reducing the amount of a serine protease or a serine protease zymogen in a plasma-derived target protein composition, the method comprising the steps of:
(a) enriching a plasma-derived target protein, in a first target protein enrichment step comprising alcohol fractionation of a Cohn pool, thereby forming a first enriched plasma-derived target protein composition
(b) contacting the first enriched plasma-derived target protein composition, or a further enriched composition thereof, with finely divided silicon dioxide (SiO 2 ) under conditions suitable to bind at least one serine protease or serine protease zymogen, the conditions comprising a conductivity of from 0.1 mS/cm to 3.0 mS/cm and a pH of from 4.0 to 7.0; and
(c) separating the SiO 2 from the enriched plasma-derived target protein composition in (b) to remove the bound serine protease,
wherein the at least one serine protease or serine protease zymogen is Factor XIa (FXIa), Factor XIIa (FXIIa), Factor XI (FXI), or Factor XII (FXII) and further wherein the plasma-derived target protein is selected from albumin, alpha-1-antitrypsin (A1PI), a protein of the complement system, and an inter-alpha-trypsin inhibitor (IαI).
2. The method of claim 1 , wherein the plasma-derived target protein is a protein of the complement system selected from the group consisting of Factor H (FH), Factor D, complement protein C3, and C4 binding protein.
3. The method of claim 1 , wherein the plasma-derived target protein composition is a manufacturing intermediate.
4. The method of claim 1 , wherein the method further comprises a second target protein enrichment step, thereby forming a second enriched plasma-derived target protein composition, after the first target protein enrichment step and prior to contacting the enriched plasma-derived target protein composition with finely divided silicon dioxide (SiO 2 ).
5. The method of claim 4 , wherein the second target protein enrichment step is a protein precipitation step.
6. The method of claim 4 , wherein the second target protein enrichment step is a chromatographic step.
7. The method of claim 4 , wherein the method further comprises a third target protein enrichment step after separating the SiO 2 from the enriched plasma-derived target protein composition to remove the bound serine protease.
8. The method of claim 7 , wherein the second target protein enrichment step is a protein precipitation step and the third target protein enrichment step is a chromatographic step.
9. The method of claim 7 , wherein the second target protein enrichment step is a chromatographic step and the third target protein enrichment step is a chromatographic step.
10. The method of claim 8 , wherein the plasma-derived target protein is Factor H.
11. The method of claim 9 , wherein the plasma-derived target protein is Factor H.
12. The method of claim 8 , wherein the plasma-derived target protein is an inter-alpha-trypsin inhibitor (IαI).
13. The method of claim 9 , wherein the plasma-derived target protein is an inter-alpha-trypsin inhibitor (IαI).
14. The method of claim 1 , wherein the composition is contacted with SiO 2 at a final concentration of from about 1 g SiO 2 /g protein to about 5 g SiO 2 /g protein.
15. The method of claim 1 , wherein the conditions suitable to bind at least one serine protease or serine protease zymogen comprise a conductivity of from 0.1 mS/cm to 2.0 mS/cm and a pH of from 4.0 to 7.0.
16. The method of claim 1 , wherein the conditions suitable to bind at least one serine protease or serine protease zymogen comprise a conductivity of from 0.5 mS/cm to 2.0 mS/cm and a pH of from 4.5 to 6.5.
17. The method of claim 1 , wherein the Cohn pool is cryo-poor plasma.
18. The method of claim 1 , wherein the alcohol is ethanol.