[Affinity chromatography for purification of phospholipase A2 from the venom of Central Asian cobra]

A method of affinity chromatography was developed for purification of phospholipase A2(PL-A2) from the Central Asian cobra venon. The enzyme was covalently coupled to a polyamide sorbent with phosphatidilethanolamine (PEA) and cytotoxin (CT). The effect of CA2+ concentration and the ion strength of the solution on the enzyme adsorption was studied. The most efficient coupling of the enzyme to the sorbent was observed at pH 8--9 in case of the Ca2+ absence and a low ion strength of the solution. For desorption of the enzyme Triton X-100 at a concentration of 0.5% should be introduced in the eluting solution. The affinity adsorption chromatography enabled the isolation of two forms of phospholipase A2 with different affinity for PEA and CT. The total yield of the enzyme was 91% at a purification degree of 5.5 and 3.5, respectively. The introduction of the second ligand (CT) in the composition of the sorbent with the phospholipid ligand allowed the authors to increase its capacity and affinity for the phospholipase A2 from the snake venom.