Immunoaffinity purification of bluetongue virus group specific antibody using recombinant protein adsorbed to polystyrene wells

Abstract

Most of the common viral diseases can be diagnosed by serological assays and these assays mostly depend on the purity and quality of antibody used. Such specific antibodies can be generated by hybridoma technology. Alternatively, the virusspecific antibodies can be purified from polyclonal serum by immunoaffinity purification technique. Based on this immune affinity purification technique we have purified group-specific antibody against Bluetongue virus (BTV) using recombinant protein VP7 of BTV bound to polystyrene wells. Elution buffer consisting of 100 mM Glycine-HCl, pH 3.0 was found optimum for dissociation of the antibody from recombinant antigen and also to maintain the integrity of antigen. The reactivity of eluted antibody was tested in enzyme-linked immunosorbent assay (ELISA). The purified antibody will be useful in other serological assays like ELISA, fluorescent assay, and agar gel immunodiffusion (AGID) for Bluetongue disease diagnosis.