Comparison of four different purification methods for isolation of anti Echis carinatus antivenom antibodies from immunized chicken egg yolk

Egg-laying hens were immunized with Echis carinatus venom and the antibodies were extracted from egg yolk by four different purification methods. The chicken egg yolk antibodies were purified by water dilution method, Polyethylene glycol and ammonium sulphate precipitation method, chloroform extraction method and Lithium sulphate precipitation method. These methods were compared in terms of total protein content, immunospecific anti Echis carinatus IgY activity, in vitro and invivo neutralizing capacity of IgY against Echis carinatus venom. Total immunoglobulin Y (IgY) concentrations varied from 1.6 to 7.0 mg per ml of egg yolk. In Neutralization studies IgY Purified by PEG and Ammonium sulphate precipitation (PEG-AS) showed better results when compared to other purification methods. About 1.25 mg of IgY (PEG-AS) able to neutralize 2LD50 Echis carinatus venom. The results indicate that the purification of IgY by PEG and Ammonium sulphate results in very pure IgY in high quantities (93% +/- 5% of total egg yolk protein) and also capable of neutralizing toxic and lethal components of Echis carinatus venom.