Abstract
In the present study, a total of 24 MAbs were produced against bluetongue virus (BTV) by polyethyleneglycol (PEG) mediated fusion method using sensitized lymphocytes and myeloma cells. All these clones were characterized for their reactivity to whole virus and recombinant BTV-VP7 protein, titres, isotypes and their reactivity with 24 BTV-serotype specific sera in cELISA. Out of 24 clones, a majority of them (n = 18) belong to various IgG subclasses and the remaining (n = 6) to the IgM class. A panel of eight clones reactive to both whole BTV and purified rVP7 protein were identified based on their reactivity in iELISA. For competitive ELISA, the clone designated as 4A10 showed better inhibition to hyperimmune serum of BTV serotype 23. However, this clone showed a variable percent of inhibition ranging from16.6% with BTV 12 serotype to 78.9% with BTV16 serotype using 24 serotype specific sera of BTV originating from guinea pig at their lowest dilutions. From the available panel of clones, only 4A10 was found to have a possible diagnostic application.
References
Anderson J. 1984. Use of monoclonal antibodies in a blocking ELISA to detect group specific antibodies to bluetongue virus. J Immunol Methods, 7(1): 139–149.
Cherrington J M, Ghalib H W, Sawyer M M, et al. 1985. Detection of viral antigens in bluetongue virus-infected ovine tissues, using the peroxidase-antiperoxidase technique. Am J Vet Res, 46: 2356–2359.
Coligan J E, Kruisbeek A M, Margulies D H, et al. 1995. In: Current protocols in immunology, Vol. 1, Chapter 2.5, Bethesda: National Institute of Health, Copyright by John Wiley & Sons, Inc. p.2.5.1–2.5.17.
Fauquet C M, Mayo M A, Maniloff J, et al. 2005. In: Virus Taxonomy. Eighth Report of the International Committee on Taxonomy of Viruses, VIII ed. Amsterdam: Elsevier-Academic Press, p515–520.
Foster N M, Luedke A J. 1968. Direct assay for bluetongue virus by intravascular inoculation of embryonating chicken eggs. Am J Vet Res, 29: 749–753.
Holbrook F R, Tabachnick W J, Schmidtmann E T, et al. 2000. Sympatry in the Culicoides variipennis complex (Diptera: Ceratopogonidae): a taxonomic reassessment. J Med Entomol, 37: 65–76.
http://www.oie.int/eng/normes/mmanual/A_00032.htm (1 of 18)12/26/2006 1:00:39 PM.
http://www.oie.int/eng/normes/mmanual/2008/pdf/2.01.03_BLUETONGUE.pdf. 08/12/2010 17: 57: 30 PM.
Jeggo M H, Gumm I D, Taylor W P. 1983. Clinical and serological response of sheep to serial challenge with different bluetongue virus types. Res Vet Sci, 34: 205–211.
Mecham J O. 1993. Detection of bluetongue virus from blood of infected sheep by use of an antigen-capture enzyme-linked immunosorbent assay after amplification of the virus in cell culture. Am J Vet Res, 54(3): 370–372.
Mertens P P C, Burroughs J N, Anderson J. 1987. Purification and properties of virus particles, infectious subviral particles, and cores of bluetongue virus serotypes 1 and 4. Virology, 157: 375–386.
Orru G, Ferrando M L, Meloni M, et al. 2006. Rapid detection and quantitation of bluetongue virus (BTV) using a molecular beacon fluorescent probe assay. J Virol Methods, 137(1): 34–42.
Pathak K B, Biswas S K, Tembhurne P A, et al. 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(1–2): 6–12.
Peters J H, Baumgarten H. 1992. Monoclonal antibodies, 1st Ed. Springer Laboratory.
Portanti O, Luciani M, Ronchi G F. 2005. Rapid detection of bluetongue virus in blood and organ samples using a capture enzyme-linked Immunosorbent assay. Vet Ital, 41(1): 15–21.
Stanislawek W, Lunt R, Blacksell S, et al. 1996. Detection by ELISA of bluetongue antigen directly in the blood of experimentally infected sheep. Vet Microbiol, 52: 1–12.
Wechsler S J, McHolland L E. 1988. Susceptibilities of 14 cell lines to bluetongue virus infection. J Clin Microbiol, 26: 2324–2327.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Bhanuprakash, V., Hosamani, M., Balamurugan, V. et al. Production and characterization of Monoclonal antibodies to bluetongue virus. Virol. Sin. 26, 8–18 (2011). https://doi.org/10.1007/s12250-011-3171-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12250-011-3171-8
Key words
- Bluetongue virus
- Competitive ELISA
- Enzyme-linked immunosorbent assay
- Monoclonal antibody
- India