Selection and characterization of single-chain recombinant antibodies against spring viraemia of carp virus from mouse phage display library

J Virol Methods. 2013 Dec;194(1-2):178-84. doi: 10.1016/j.jviromet.2013.08.017. Epub 2013 Aug 29.

Abstract

Antibody-displaying phage library was selected after three rounds of panning against spring viraemia of carp virus (SVCV) by phage display technology. Eight positive clones which could produce soluble single-chain fragment variable (scFv) antibody induced by isopropyl-beta-d-thiogalactopyranoside (IPTG) were obtained. Dot blot results showed that the eight scFv antibodies could recognize SVCV. The soluble scFv antibodies showed a molecular weight 29 kD by Western blot. All scFv antibodies could recognize SVCV proteins specifically without cross-reaction with other virus proteins by ELISA. Indirect immunofluorescence results showed that all of these scFv antibodies reacted positively with virus in the SVCV-infected cells. These scFv antibodies will be useful tools to establish immunological detection methods for SVCV.

Keywords: Phage display; Single-chain fragment variable; Spring viraemia of carp virus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Viral / chemistry
  • Antibodies, Viral / immunology*
  • Antibodies, Viral / isolation & purification*
  • Blotting, Western
  • Carps
  • Cell Surface Display Techniques
  • Mice
  • Molecular Weight
  • Single-Chain Antibodies / chemistry
  • Single-Chain Antibodies / immunology*
  • Single-Chain Antibodies / isolation & purification*
  • Vesiculovirus / immunology*

Substances

  • Antibodies, Viral
  • Single-Chain Antibodies