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VirusDisease

, Volume 27, Issue 1, pp 98–101 | Cite as

Purification of infective bluetongue virus particles by immuno-affinity chromatography using anti-core antibody

  • Karam Chand
  • Sanchay K. Biswas
  • Bimalendu MondalEmail author
Short Communication

Abstract

An immuno-affinity chromatography technique for purification of infective bluetongue virus (BTV) has been descried using anti-core antibodies. BTV anti-core antibodies (prepared in guinea pig) were mixed with cell culture-grown BTV-1 and then the mixture was added to the cyanogens bromide-activated protein-A Sepharose column. Protein A binds to the antibody which in turn binds to the antigen (i.e. BTV). After thorough washing, antigen–antibody and antibody-protein A couplings were dissociated with 4M MgCl2, pH6.5. Antibody molecules were removed by dialysis and virus particles were concentrated by spin column ultrafiltration. Dialyzed and concentrated material was tested positive for BTV antigen by a sandwich ELISA and the infectivity of the chromatography-purified virus was demonstrated in cell culture. This method was applied for selective capture of BTV from a mixture of other viruses. As group-specific antibodies (against BTV core) were used to capture the virus, it is expected that virus of all BTV serotypes could be purified by this method. This method will be helpful for selective capture and enrichment of BTV from concurrently infected blood or tissue samples for efficient isolation in cell culture. Further, this method can be used for small scale purification of BTV avoiding ultracentrifugation.

Keywords

Anti-core antibody Bluetongue virus Immuno-affinity chromatography 

Notes

Acknowledgments

This work has partly been supported by an ICAR funded project—All India Network Program on Bluetongue. Authors are grateful to the Director and Head Virology Division of IVRI for providing facilities to carry out this work.

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Copyright information

© Indian Virological Society 2016

Authors and Affiliations

  • Karam Chand
    • 1
  • Sanchay K. Biswas
    • 1
  • Bimalendu Mondal
    • 1
    • 2
    Email author
  1. 1.Division of VirologyIndian Veterinary Research InstituteDistrict NainitalIndia
  2. 2.Eastern Regional StationIndian Veterinary Research InstituteKolkataIndia

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