Purification and characterization of an antihemorrhagic factor in the serum of Trimeresurus flavoviridis, a crotalid

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1. 1. An antihemorrhagic factor in the serum of Trimeresurus flavoviridis, a crotalid, was purified by (NH 4) 2SO 4 fractionation, ethanol fractionation, gel filtration on Sephadex G-200, chromatography on DEAE-cellulose and isoelectric focusing in carrier ampholytes. A 21-fold increase in specific activity was attained with a yield of 15%. The purified preparation was homogenous as judged by ultracentrifugation, gel filtration on Sephadex G-200, disc electrophoresis, immunoelectrophoresis and isoelectric focusing. 2. 2. The purified serum factor inhibited the two immunologically distinct hemorhagic principles, HR1 and HR2 (mixture of HR2a and 2b), in the venom of this snake and antihemorrhagic activities against HR1 and HR2 increased simultaneously during purification. The purified factor inhibited not only the hemorrhagic activities but also the lethal toxicity in the venom. 3. 3. The molecular weight of the purified factor was approx. 70 000 ( s° 20, w = 4.05); the isoelectric point was around 4. The purified factor migrated to a position in the area of albumin and α 1-globulin in immunoelectrophoresis and did not form any precipitin line with the crude venom or with the purified hemorrhagic principles in immunodiffusion tests. The relation of the serum factor to immunoglobulins was discussed. 4. 4. The crude serum from T. flavoviridis inhibited hemorrhagic activities of all snake venoms tested, including Crotalidae, Viperidae and Elapidae venoms. A possible mechanism of the inhibition by the serum factor of the hemorrhagic activity of snake venoms was also discussed.