Indian Journal of Animal Research

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Indian Journal of Animal Research, volume 52 issue 1 (january 2018) : 126-130

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

Karam Chand, Sanchay K. Biswas, Bimalendu Mondal, Awadh B.Pandey
1Division of Virology, Indian Veterinary Research Institute, Mukteswar Campus, Dist. Nainital-263 138, Uttarakhand, India
Cite article:- Chand Karam, Biswas K. Sanchay, Mondal Bimalendu, B.Pandey Awadh (2017). Immunoaffinity purification of bluetongue virus group specific antibody using recombinant protein adsorbed to polystyrene wells. Indian Journal of Animal Research. 52(1): 126-130. doi: 10.18805/ijar.v0iOF.9137.

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 virus-specific 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.

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