Fractionation of Bee Venom I Preparation and Characterization of Four Antigenic Components

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Abstract
A tandem column system packed with Sephadex G-50 and Sephadex G-75 in 0.1 M NH4CHO2 pH 4.5 has been used to fractionate bee venom. The results of the fractionation show that bee venom is composed of 8 or more components. Rechromatography of all single components to remove contamination by other components, has been done. Bio-Gel E-150 and P-2 columns, as well as Sephadex G-25, G-50 and G-75 columns were used. A Sephadex G-50 column eluted with buffer plus 8M Urea was used to purify melittin. This was followed, after desalting on Bio-Gel P-10, by ion exchange chromatography on Sephadex CM-25. Phospholipase, hyaluronidase, melittin, apamin, and vanilmandelic acid were located in the chromatogram. Purity of four components was shown by immunoelectrophoresis.