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 preparing an Immunoglobulin G (IgG) composition having a reduced amount of Factor XIa, the method comprising the steps of:
(a) precipitating a cryo-poor plasma fraction, in a first precipitation step, with from about 6% to about 10% alcohol at a pH of from about 7.0 to about 7.5 to obtain a first precipitate and a first supernatant;
(b) precipitating IgG from the first supernatant, in a second precipitation step, with from about 23% to about 27% alcohol at a pH of from about 6.7 to about 7.3 to form a second precipitate;
(c) suspending the second precipitate to form a first suspension;
(d) contacting the first suspension with fumed silica under a solution condition suitable to bind Factor XIa; and
(e) separating the fumed silica from the first suspension to form a clarified suspension, thereby forming an IgG composition having a reduced amount of Factor XIa.
2. The method of claim 1 , wherein the method further comprises the steps of:
(f) precipitating IgG from the clarified suspension formed in step (e), in a third precipitation step, with from about 22% to about 28% alcohol at a pH of from about 6.7 to about 7.3 to form a third precipitate;
(g) suspending the third precipitate to form a second suspension; and
(h) separating a soluble fraction from an insoluble fraction in the second suspension formed in step (g), thereby forming an enriched IgG composition.
3. The method of claim 1 , wherein the method further comprises an anion exchange chromatography enrichment step.
4. The method of claim 1 , wherein the method further comprises a cation exchange chromatography enrichment step.
5. The method of claim 1 , wherein the method further comprises at least one dedicated viral inactivation or removal step.
6. The method of claim 5 , wherein the method comprises a solvent/detergent (S/D) viral inactivation step.
7. The method of claim 5 , wherein the method comprises a nanofiltration step.
8. The method of claim 5 , wherein the method comprises an incubation step at low pH.
9. The method of claim 1 , wherein step (b) comprises adjusting the ethanol concentration of the first supernatant formed in step (a) to about 25% (v/v) at a temperature from about −7° C. to about −9° C.
10. The method of claim 1 , wherein step (d) comprises the addition fumed silica to a final concentration of from about 0.02 grams per gram precipitate formed in step (b) to about 0.06 grams per gram precipitate formed in step (b).
11. The method of claim 1 , wherein the solution condition suitable to bind Factor XIa comprises a pH of from 4.5 to 6.0 and a conductivity of from 0.1 mS/cm to 3 mS/cm.
12. The method of claim 1 , wherein the first suspension is contacted with fumed silica at a pH of from 4.9 to 5.3.
13. The method of claim 12 , wherein the first suspension is contacted with fumed silica at a conductivity of from 0.5 mS/cm to 2 mS/cm.
14. The method of claim 1 , further comprising the sub-step of:
(iv) treating the clarified suspension with a detergent.
15. The method of claim 2 , wherein the enriched IgG composition obtained in step (h) contains at least 85% of the IgG content found in the cryo-poor plasma fraction used in step (a).
16. The method of claim 15 , wherein the enriched IgG composition obtained in step (h) contains at least 90% of the IgG content found in the cryo-poor plasma fraction used in step (a).
17. The method of claim 2 , wherein step (b) comprises adjusting the ethanol concentration of the first supernatant formed in the step (a) to about 25% (v/v) at a temperature of about −7° C. to about −9° C.
18. The method of claim 1 , wherein the first suspension is contacted with fumed silica at a pH of from 4.9 to 5.3 and a conductivity of from 0.5 mS/cm to 2 mS/cm.