A HIGH-EFFICIENCY METHOD FOR PURIFICATION AND ASSAY OF BEE VENOM PHOSPHOLIPASE-A2

Phospholipase A(2) (PLA(2)) is the main allergen of hymenopteran venom s. We describe a highly efficient reverse phase high performance liqui d chromatographic method (HPLC) for isolating PLA(2) from crude bee ve nom. This method removes all detectable contaminants such as melittin from PLA(2) while preserving the hemolytic action of PLA(2). In additi on we describe a simple functional assay of PLA(2) based on its propen sity to cause hemolysis of guina pig red blood cells. These techniques are particularly well suited to the isolation and assessment of PLA(2 ) of venoms which are available in limited quantities.