Production of monoclonal antibodies binding to the VP7 protein of African horse sickness virus

J Virol Methods. 2019 Aug:270:131-136. doi: 10.1016/j.jviromet.2019.05.004. Epub 2019 May 13.

Abstract

Monoclonal antibodies (MAbs) against AHSV were produced by immunising BALB/c mice with AHSV serotype 9 and six clones able to recognize specifically the VP7-AHSV with a strong reactivity were selected. The specificity of the MAbs was assessed in i-ELISA against a commercial VP7-AHSV and in immunoblot against a home-made VP7-AHSV, expressed by a Baculovirus expression system; potential cross-reactions with related orbiviruses (Bluetongue virus and Epizootic Haemorrhagic Disease virus) were investigated as well. One of the six MAbs selected, MAb 7F11E14, was tested in direct immunofluorescence and reacted with all nine AHSV serotypes, but didn't cross-react with BTV and EHDV. MAb 7F11E14 was also used to develop a competitive ELISA and was able to detect AHSV antibodies in the sera of AHS infected animals.

Keywords: African horse sickness; ELISA; Immunofluorescence; Monoclonal antibodies.

MeSH terms

  • African Horse Sickness / diagnosis*
  • African Horse Sickness / immunology*
  • African Horse Sickness Virus / immunology*
  • Animals
  • Antibodies, Monoclonal / blood*
  • Antibodies, Viral / blood
  • Antigens, Viral / immunology
  • Bluetongue virus / immunology
  • Cross Reactions
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Hemorrhagic Disease Virus, Epizootic / immunology
  • Horses
  • Mice
  • Mice, Inbred BALB C
  • Recombinant Proteins
  • Sensitivity and Specificity
  • Viral Core Proteins / immunology*
  • Viral Core Proteins / isolation & purification

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Antigens, Viral
  • Recombinant Proteins
  • Viral Core Proteins