SIMULTANEOUS ISOLATION OF PROTEIN-C ACTIVATOR, FIBRIN CLOT PROMOTING ENZYME (FIPROZYME) AND PHOSPHOLIPASE A(2) FROM THE VENOM OF THE SOUTHERN COPPERHEAD SNAKE

The simultaneous isolation of three enzymes from the southern copperhe ad snake venom (Agkistrodon contortrix contortrix; ACC) is described. The first step is a chromatography of crude venom on a Mono S cation-e xchange column at pH 6.5. A fibrin clot promoting enzyme (fiprozyme) t hat preferentially releases fibrinopeptide B from fibrinogen is isolat ed from the fraction not binding to the Mono S by a further three-step process. The procedure involves affinity chromatography on Blue Sepha rose, gel chromatography on Sephacryl S-200 and metal-chelate chromato graphy on Chelating Sepharose. Protein C activator and phospholipase c oelute from the Mono S column. They are separated by a gel chromatogra phy on Sephacryl S-200. After this step two enzymes are obtained: a hi ghly purified protein C activator applicable in methods for determinat ion of functional level of protein C (a plasma regulator of hemostasis ) and an electrophoretically pure enzyme with the activity of phosphol ipase A(2).