Compounds and methods for purification of serine proteases
View Patent ↗Disclosed herein are compounds, compositions, methods and kits for purifying a serine protease and serine proteases purified with the compounds, compositions and methods.
1. A compound selected from:
or a salt thereof.
2. An affinity solid support selected from:
or a salt thereof,
wherein:
Y—Z is
Z is a solid support;
q is 3, 4 or 6; and
v is 1, 2, 3, 4, 5, 6 or 7.
3. The affinity solid support of claim 2 , wherein Z is an agarose resin.
4. A kit for purifying a serine protease comprising
(1) an affinity solid support of claim 2 , and
(2) an elution buffer comprising a competitive agent.
5. A kit for purifying a serine protease comprising a compound of claim 1 and a resin of the formula NC—O—Z or
wherein Z is a solid support.
6. The kit of claim 5 , wherein Z is an agarose resin.
7. A method for purifying a fXa derivative comprising
(1) adding a first composition comprising the fXa derivative to an affinity solid support of claim 2 or a salt thereof to form a second composition comprising the fXa derivative and the affinity solid support of claim 2 , and
(2) eluting the fXa derivative from the second composition with an elution buffer comprising a competitive agent,
wherein the fXa derivative is a polypeptide comprising the amino acid sequence of SEQ ID NO: 1, 2 or 4, or a polypeptide having at least about 80% sequence identity to SEQ ID NO: 1, 2 or 4 and further wherein the derivative is capable of binding to a fXa inhibitor and does not assemble into a prothrombinase complex.
8. The method of claim 7 , wherein the competitive agent is arginine and/or benzamidine, or a salt thereof.
9. The method of claim 7 , wherein the fXa derivative is a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, or a polypeptide having at least about 90% sequence identity to SEQ ID NO: 2.
10. The method of claim 9 , wherein the polypeptide has at least about 95% sequence identity to SEQ ID NO: 2.