An improved method for isolation of anti-viper venom antibodies from chicken egg yolk

The production of antibodies and its purification from mammalian blood has been found low yielding and laborious. Therefore, anti snake venom antibodies for therapeutic use is obtained mostly as polyvalent whole serum or partially purified polyvalent immunoglobulin. The side effects of anti snake venom (ASV) therapy are mainly serum sickness and renal failure, which may be reduced by using sufficiently pure antibodies. Therefore, we have standardized a simple method for production of purified antivenom. Here, we present the development of polyclonal antibodies against viper venom in hens and its isolation from the egg yolk of immunized birds. We have modified the reported methods of purification of immunoglobulin from egg yolk, and thus yielded 90% purity of the protein. The modified method involves only two steps, such as removal of lipids from the diluted egg yolk by a freeze-thaw cycle and centrifugation, followed by gel filtration on Biogel P-150. The advantages are that the process is very simple, and from one egg, 100+/-20 mg of pure immunoglobulin is obtained. The antibodies are present in the egg for up to 100 days after the immunization. Thus, using small amounts of venom, a large quantity of the immunoglobulin is obtained in a sufficiently pure form. The antigen binding ability of the pure antibody is found good by the Ouchterlony's double diffusion experiment.