Rapid purification of high purity thrombin and preparation of a novel hemostat for clinical purposes

Abstract

Thrombin was prepared from crude prothrombin enriched plasma by activation using Russell's viper venom. Prothrombin was prepared by barium sulphate adsorption and elution of prothrombin enriched fraction using high concentrations of sodium citrate. This fraction was directly activated with venom and thrombin was purifi ed by SP Sepharose ion-exchange chromatography and subsequently over Phenyl-sepharose column. This product exhibits a purity of >98% with an activity of at least 6000U/ mg or higher. The Thrombin was further used in the preparation of a novel bio-absorbable hemostat using Calcium Alginate fi ber sheet which acts as an absorbable hemostat. The present hemostat is highly porous, easy to use and has faster clotting time due to higher solubility of calcium fi bers and thereby releasing thrombin.