Production and assessment of polyclonal antibody against the whole virion of Goatpox virus and sheeppox virus in the rabbit

Abstract

Background: Antibodies have emerged as essential tools of biomedical researches and are of great commercial and medical values. They are the fastest growing product segments of the pharmaceutical industry. Polyclonal antibodies are antibodies that secreted by different B cell lineages within the body. In this study production of polyclonal antibody against Goatpox and Sheeppox virion in rabbit was performed and subsequently; it will be used as the development of ELISA for detection of poxvirus in different species of animals. Results: After 0.5mg/ml of the whole virion of the poxvirus was injected subcutaneously at multiple sites, the blood was collected at an interval of 14 days and antibody titration was conducted. GTPV A27 antigen was coated and indirect ELISA method was used for the titration of antibody then the mean of OD values for positive and negative results were analyzed by Microsoft excel window 7. The blood was collected and serum was prepared for IgG purification. The IgG was purified by Ammonium sulfate precipitation method which was dialyzed against 5lire PBS overnight at +4oc. The protein concentration was determined spectrophotometrically at 280nm wavelength and estimated as 2.29µg/µl and 2.18µg/µl against Goatpox virus and sheeppox virus respectively. Protein bands were checked by SDS PAGE and the molecular weights of 67kDa and 25kDas were estimated for IgG. similarly, antigen-antibody detection was checked by western blot. Conclusions: Based on the results of this study, the virion of Goatpox virus and Sheeppox virus is strong enough to produce antibody to develop ELISA kit for detection of poxviruses by coating GPTV A27 antigen. This study is the first to be conducted for the production of antibody from virions of Goatpox and sheeppox virus which was tittered by coating GTPV A27 antigen.