Indian Journal of Animal Research

  • Chief EditorK.M.L. Pathak

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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.
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.
  1. Afshar, A., Thomas, F.C., Wright, P.F., Shapiro, J.L. and Anderson, J. (1989). Comparison of competitive ELISA, indirect ELISA and standard AGID tests for detecting blue­tongue virus antibodies in cattle and sheep. Vet Rec. 124: 136-141.
  2. Bayry,J., Prabhudas, K., Bist, P., Reddy, G. R. and Suryanarayana, V.V.S. (1999). Immuno affinity purification of foot and mouth disease virus type specific antibodies using recombinant protein adsorbed to polystyrene wells. J. Virol. Methods. 81: 21-30
  3. Belhouchet, M., MohdJaafar, F., Firth, A.E., Grimes, J.M., Mertens, P. P. and Attoui, H. (2011). Detection of a fourth orbivirus non-    structural protein. PLoS one, 6:e25697.
  4. Brahimi, K., Perignon, J.L., Bossus, M.., Gras, H., Tartar, A. and Druilhe, P. (1993). Fast immunopurification of small amounts of specific antibodies bound to ELISA plates. J. Immunol. Methods. 162: 69-75.
  5. Chand, K., Biswas, S.K., De, A., Sing, B. and Mondal, B. (2009). A polyclonal antibody-based sandwich ELISA for the detection of bluetongue virus in cell culture and blood of sheep infected experimentally. J Virol Methods. 160:189-92.
  6. Chand, K., Biswas, S.K., Pandey, A.B., Muthuchelvan, D. and Mondal, B. (2015). Bluetongue in India: A review. Adv Anim Vet Sci. 3(11):605-612.
  7. Hussein el , A.B., Calisher, C.H., Holbrook, F.R., Schoepp, R.J. and Beaty, B.J. (1989) Detection of bluetongue virus antigens in Culicoides variipennis by enzyme immunoassay. J Clin Microbiol. 27(6):1320-1323.
  8. Huismans, H. and Erasmus, B.J. (1981). Identification of the serotype-specific and group-specific antigens of blue­tongue virus. Onderstepoort J Vet. 48: 51-58.
  9. Jun Gu, Stephenson, C.G. and Ladarola, M.J. (1994). Recombinant proteins attached to a Nickel column: Use in affinity purification of antibodies. Biotechniques. 17: 257-262.
  10. Mallorquí, J., Llop, E., De Bolos, C., Gutiérrez-Gallego, R., Segura, J. and Pascual, J.A. (2010). Purification of erythropoietin from human plasma samples using an immunoaffinity well plate. J Chromatogr B. 878(23): 2117-2122.
  11. Mertens, P.P.C., Maan, S., Samuel, A. and Attoui, H. (2004). Orbivirus, Reoviridae. In Virus Taxonomy, VIIIth Report of the ICTV, pp. 466-483. Edited by C.M. Fauquet, M.A. Mayo, J. Maniloff, U. Desselberegr and L.A. Ball. Elsevier/ Academic Press, London. 
  12. Pathak, K. B., Biswas, S. K., Tembhurne, P. A., Hosamani, M., Bhanuprakash, V. Prasad, G., Singh, R. K., Rasool, T. J. and Mondal, B. (2008). Prokaryotic expression of truncated VP7 of Bluetongue virus (BTV) and reactivity of the purified recombinant protein with all BTV type-specific sera. J Virol Methods. 152: 6-12.
  13. Pullen, G.R., Fitzgerald, M.G. and Hosking, C.S. (1986). Antibody avidity determination by ELISA using thiocyanate elution. J Limmunol Methods, 86(1): 83-87
  14. Xu, R., Lin, G., Wang, W., Liu, M., Zhan, S., Wang, L., Zhang, K., Zhang, R. and Li, J. (2010). Application of an ELISA elution assay to dissociate digoxin antibody complexes in immunoaffinity chromatography. Scand J Immunol. 71(1):55-60. 
  15. Zientara, S., Sailleau, C., Viarouge, C., Hoper, D., Beer, M., Jenckel, M., Hoffmann, B., Romey, A., Bakkali-Kassimi, L., A. and Vitour, Dl. (2014). Novel bluetongue virus in goats, Corsica, France, Emerg Infect Dis. 20: 2123-2125 

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