Bovine thrombin purification

摘要

Disclosed is a method for the preparation of a purified bovine thrombin composition, substantially free of impurities having a molecular weight greater than 40 kDa, and having a specific activity greater than 1800 units of thrombin per mg (u/mg) of protein, which method comprises: (a) applying a bovine thrombin preparation having from about 500 to about 3000 units of thrombin per mL (u/mL) of the preparation to a cation exchange chromatography to afford a pre-purified bovine thrombin composition having a specific activity greater than about 1500 u/mg of protein; (b) applying the pre-purified bovine thrombin composition from step (a) to a size exclusion filtration capable of excluding impurities having a molecular weight greater than 40 kDa to afford a purified bovine thrombin composition, substantially free of impurities having a molecular weight greater than 40 kDa, and having a specific activity greater than 1800 u/mg of protein; and optionally, (c) applying the purified bovine thrombin composition from step (b) to an anion exchange filtration.