Methods for processing coagulation factors

摘要

The present application relates to methods for purifying recombinant coagulation factor proteins, including for example, prothrombin (Factor II). In embodiments, the methods provide purified prothrombin that exhibit increased bioactivity and reduced levels of thrombin, thereby increasing the safety of the prothrombin. Also provided are purified recombinant coagulation factor proteins.

主权项

1. A method for purifying a recombinant coagulation factor protein, the method comprising: a) providing a mixture comprising the recombinant coagulation factor protein; b) filtering the mixture through at least a first anion exchange column to produce a first anion exchange column product; c) loading the first anion exchange column product onto a poly(ethyleneimine) (PEI) column and eluting with a linear sodium chloride gradient to produce one or more fractions of a PEI column product; and d) recovering one or more fractions of PEI column product with a combined pre-peak of less than 30% to obtain the purified recombinant coagulation factor protein, wherein the purified recombinant coagulation factor protein comprises less than about 0.8/10, 0.7/10, 0.6/10, 0.5/10 or 0.4/10 missing D-carboxylated glutamic acid sites and wherein the pre-peak of one or more PEI column product is determined by high performance liquid chromatography (HPLC).