Factor X activation

Patent metadata

Field	Value
Publication number	US-10036002-B2
Application number	US-201514824870-A
Country	US
Kind code	B2
Filing date	Aug 12, 2015
Priority date	Aug 12, 2014
Publication date	Jul 31, 2018
Grant date	Jul 31, 2018

Abstract

Methods and systems for activating Factor X are disclosed.

First claim

The invention claimed is:

1. A method for activating Coagulation Factor X (FX) comprising:
a) providing an aqueous solution that comprises FX isolated from mammalian plasma or tissue culture cell supernatant;
b) contacting the aqueous solution with an anion-exchange material under binding conditions for a contact period of at least five hours; and
c) eluting the activated FX (FXa) from the anion-exchange material with an elution buffer.

2. The method of claim 1 , wherein the contact period is at least 10 hours.

3. The method of claim 1 , wherein the binding conditions comprise a conductivity of 8 mS/cm or less.

4. The method of claim 1 , wherein the binding conditions comprise a pH of between 8 and 10.

5. The method of claim 1 , wherein the FX is loaded at greater than 0.5 mg per mL of anion-exchange material.

6. The method of claim 1 , wherein the contact and eluting steps are performed at a temperature of greater than 8° C.

7. The method of claim 1 , wherein the elution buffer comprises at least one of a divalent cation and a counter-ion.

8. The method of claim 7 , wherein the divalent cation is Ca 2+ .

9. The method of claim 7 , wherein the counter-ion is at least one of chloride, acetate, or sulfate.

10. The method of claim 7 , wherein the concentration of the divalent cation or counter-anion or both in the elution buffer increases over time.

11. The method of claim 10 , wherein the concentration of the divalent cation increases from 1 to 100 mM.

12. The method of claim 10 , wherein the concentration of the divalent cation increases from 1 to 10 mM.

13. The method of claim 10 , wherein the concentration of the counter-anion increases from 1 to 1000 mM.

14. The method of claim 10 , wherein the concentration of the counter-anion increases from 9 to 200 mM.

15. The method of claim 1 , wherein the elution buffer has a conductivity between 1 and 100 mS/cm at 25° C.

16. The method of claim 1 , wherein the aqueous solution comprising FX is contacted with the anion-exchange material in the presence of ethylenediaminetetraacetic acid (EDTA).

17. The method of claim 1 , wherein the anion-exchange material comprises a ligand comprising at least one of quaternary ammonium [Q], diethylaminoethyl [DEAE], diethylaminopropyl [ANX], or primary amine.

18. The method claim 1 , wherein the anion-exchange material comprises a matrix derived from at least one of polystyrene, polymethylmetaacrylate, polyvinylbenzene, polyvinyl pyridine, cross-linked poly(styrene-divinylbenzene), sepharose, and cross linked agarose.

19. The method of claim 1 , further comprising, prior to step a), isolating FX from mammalian plasma to produce the aqueous solution.

20. The method of claim 1 , further comprising, prior to step a), isolating FX from tissue culture cell supernatant to produce aqueous solution.

21. The method of claim 1 , wherein the FX in the aqueous solution is isolated by precipitation, ultrafiltration, or chromatography.

Assignees

• Baxalta Inc
• Baxalta GmbH

Inventors

• Hasslacher Meinhard
• Gatternig Thomas
• Fiedler Christian
• Böhm Ernst
• Dockal Michael
• Horling Franziska

Classifications

• C12Y304/21006

  Coagulation factor Xa (3.4.21.6) · CPC title

• C12N9/6432 Primary

  Coagulation factor Xa (3.4.21.6) · CPC title