A method to purify cobra venom factor (CVF) from cobra venom by sequential column chromatography is described which yields a product virtually free of phospholipase A2, a common contaminant of CVF preparations. The separation of phospholipase A2 from CVF was achieved by chromatography on Cibacron blue-agarose, a resin that tightly binds cobra venom phospholipase A2. A rapid and simple hemolytic assay for the qualitative and quantitative determination of CVF based on its ability to induce bystander lysis of erythrocytes has been devised. CVF was isolated from the venom of the Naja naja kaouthia subspecies and some of its biochemical properties and physicochemical parameters were delineated.
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