Removal of serine proteases by treatment with finely divided silicon dioxide

Patent metadata

Field	Value
Publication number	US-9468675-B2
Application number	US-201414296319-A
Country	US
Kind code	B2
Filing date	Jun 4, 2014
Priority date	May 26, 2010
Publication date	Oct 18, 2016
Grant date	Oct 18, 2016

Abstract

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.

First claim

What is claimed is:

1. A method for preparing an inter-alpha-trypsin inhibitor (IαI) composition having a reduced amount of a serine protease or a serine protease zymogen, the method comprising the steps of:
(a) contacting a blood-plasma derived solution containing IαI and at least one serine protease with from 0.01 grams to 10 grams finely divided silicon dioxide (SiO 2 ) per gram protein under conditions suitable to bind the IαI and at least one serine protease, the conditions including a pH of from 4.0 to 7.0 and a conductivity of from 0.1 mS/cm to 2.0 mS/cm;
(b) separating the SiO 2 from the composition;
(c) eluting the serine protease or serine protease zymogen from the SiO 2 under conditions in which at least 50% of the IαI remains bound, the conditions including a pH of from 5.0 to 7.0 and a conductivity of less than 5 mS/cm; and
(d) eluting the IαI from the SiO 2 under conditions including a pH of from 7.0 to 8.0 and a conductivity of at least 2 mS/cm.

2. A method for preparing an inter-alpha-trypsin inhibitor (IαI) composition having a reduced amount of a serine protease or a serine protease zymogen, the method comprising the steps of:
(a) contacting a blood-plasma derived solution containing IαI and at least one serine protease with from 0.01 grams to 10 grams finely divided silicon dioxide (SiO 2 ) per gram protein under conditions suitable to bind the IαI and at least one serine protease, the conditions including a pH of from 4.0 to 7.0 and a conductivity of from 0.1 mS/cm to 2.0 mS/cm;
(b) separating the SiO 2 from the composition; and
(c) eluting the IαI from the SiO 2 under conditions in which at least 50% of the serine protease or serine protease zymogen remains bound to the SiO 2 , the conditions including a pH of from 7.0 to 8.0 and a conductivity of no more than 10 mS/cm.

3. A method for preparing an inter-alpha-trypsin inhibitor (IαI) composition having a reduced amount of a serine protease or a serine protease zymogen, the method comprising the steps of:
(a) contacting a blood-plasma derived solution containing IαI and at least one serine protease with from 0.01 grams to 10 grams finely divided silicon dioxide (SiO 2 ) per gram protein under conditions suitable to bind the IαI but not at least 50% of the at least one serine protease or serine protease zymogen, the conditions including a pH of from 7.0 to 9.0 and a conductivity of at least 10 mS/cm;
(b) separating the SiO 2 from the composition; and
(c) eluting the IαI from the SiO 2 under conditions including a pH of from 7.0 to 9.0 and a conductivity of less than 10 mS/cm.

4. A method for preparing an inter-alpha-trypsin inhibitor (IαI) composition having a reduced amount of a serine protease or a serine protease zymogen, the method comprising the steps of:
(a) contacting a blood-plasma derived solution containing inter-alpha-trypsin inhibitor (IαI) and at least one serine protease with from 0.01 grams to 10 grams finely divided silicon dioxide (SiO 2 ) per gram protein under conditions suitable to bind the serine protease or serine protease zymogen but not at least 50% of the inter-alpha-trypsin inhibitor (IαI), the conditions including a pH of from 7.0 to 9.0 and a conductivity of less than 10 mS/cm; and
(b) separating the SiO 2 from the composition.

5. The method of claim 1 , wherein the composition is contacted with SiO 2 at a final concentration of at least 1 g SiO 2 /g protein.

6. The method of claim 5 , wherein the composition is contacted with SiO 2 at a final concentration of at least 2 g SiO 2 /g protein.

7. The method of claim 5 , wherein the composition is contacted with SiO 2 at a final concentration of at least 2.5 g SiO 2 /g protein.

8. The method of claim 1 , wherein the serine protease or serine protease zymogen is Factor XI.

9. The method of claim 1 , wherein the serine protease or serine protease zymogen is Factor XII.

10. The method of claim 1 , wherein the serine protease or serine protease zymogen is Factor XIa.

11. The method of claim 1 , wherein the serine protease or serine protease zymogen is Factor XIIa.

12. The method of claim 1 , wherein the finely divided silicon dioxide (SiO 2 ) is fumed silica.

13. The method of claim 1 , wherein the conditions in step (c) include a pH of from 6.0±0.5.

14. The method of claim 1 , wherein the conditions in step (c) include a pH of rom 6.0±0.2.

15. The method of claim 2 , wherein the conditions in step (c) include a conductivity of about 6.0.

16. The method of claim 3 , wherein the conditions in step (a) include a conductivity of at least 20 mS/cm.

17. The method of claim 4 , wherein the conditions in step (a) include a conductivity of about 6.0.

18. The method of claim 2 , wherein the finely divided silicon dioxide (SiO 2 ) is fumed silica.

19. The method of claim 3 , wherein the finely divided silicon dioxide (SiO 2 ) is fumed silica.

20. The method of claim 4 , wherein the finely divided silicon dioxide (SiO 2 ) is fumed silica.

Assignees

• Baxalta Inc
• Baxalta GmbH

Inventors

• Teschner Wolfgang
• Schwarz Hans-Peter
• Madlener Ruth
• Svatos Sonja
• Pljevljakovic Azra
• Weber Alfred

Classifications

• C07K1/30

  by precipitation · CPC title

• C07K16/065 Primary

  Purification, fragmentation · CPC title

• A61K47/183

  Amino acids, e.g. glycine, EDTA or aspartame · CPC title

• B01D15/362

  Cation-exchange · CPC title

• A61K9/08

  Solutions {(composition of solutions A61K47/00)} · CPC title