Snake venom: their purification and isolation and analysis of their anti-venom anticoagulant properties

ABSTRACT

Viperidae venoms contain toxins that are direct or indirect anticoagulants that inhibit the clotting pathway, therefore increasing the risk of bleeding. Several venoms from the families Viperidae contain proteolytic enzymes that exercise some effect on the blood coagulation process. Snake venom toxins which delay blood coagulation are proteins or glycoprotein with molecular weights ranging from 6 kDa to 350 kDa. These factors inhibit blood coagulation by different mechanisms.Some snake venoms contain toxins that are direct or indirect anticoagulants, which inhibit the clotting process, thus increasing the risk of bleeding. Snake venom toxins that prolong blood coagulation are proteins or glycoproteins with molecular masses ranging from 6 to 350 kDa. The crude venom of E. carinatuswas obtained from the Venomous Animals from department of microbiology Hazara University, Mansehra [Pakistan]. Sephadex G-75 and DEAE-Sepharose columns were purchased from Pharmacia [Sweden]. CaCl2 and PT kit was purchased from Fisher Diagnostics [USA]. Protein markers were obtained from BioRad [Hercules, USA]. Other reagents and chemicals were of analytical grade from Fluka and Merck. The anticoagulant fractions [F2C and F2D] isolated in the present work were characterized as proteases, since a photolytic effect was observed on casein, BAPNA or human plasma. Our results showed that the PT value significantly increased in the F2C and F2D fractions as compared with PT value of the crude venom. Inthe present study, the venom of Echiscarinatuswas fractionated by chromatography and each fraction evaluated by PT test. These fractions showed enzymatic activity. Their main components were proteins of molecular weights of about 42, 50 and 79 kDa.. Further studies are needed to verify this hypothesis: