Affinity chromatography of phospholipase A2 from Naja naja naja (Indian cobra) venom

A rapid and improved purification procedure is described for phospholipase A2 from the Indian cobra, Naja naja naja. The procedure is based on affinity chromatography of the venom through Affi-Gel Blue to obtain a 9-fold purification in one step. However, as there are multiple forms of the enzyme in the venom and other proteins do bind to Affi-Gel Blue, further purification is achieved by DE-11 cellulose. Finally, a minor contaminant is removed by Sulfopropyl Sephadex C-25 chromatography. The resulting product was found to be pure by analytical isoelectric focusing and double diffusion precipitin tests. An isoelectric focusing column run with venom collected from a single snake gave a similar profile to that obtained with venom pooled from several snakes.