Anti-Echis carinatus venom antibodies from chicken egg yolk: isolation, purification and neutralization efficacy

High titer antibodies (IgY) were raised in egg yolk of white leghorn chicken (Gallus gallus domesticus) by immunizing with the venom of Echis carinatus (Saw scaled viper or carpet viper), an Indian venomous snake belonging to the family Viperidae. The anti-snake venom antibodies (antivenom) were isolated from egg yolk by the water dilution method, enriched by 19% sodium sulfate precipitation and purified by immunoaffinity chromatography. A single, electrophoretically pure IgY band of 180-200 kDa was obtained on SDS-PAGE. Immunoblot analysis revealed not only the specific binding of the antivenom but also dose-dependent blocking of antivenom by venom proteins. In neutralization studies, a preincubated mixture of both affinity-purified (50 mg/kg body weight) as well as partially purified (210 mg/kg body weight) anti-E. carinatus IgY with 2 LD(50) dose of E. carinatus venom (2 x 6.65 mg/kg body weight) gave 100% protection in mice when administered subcutaneously.