Eight novel single chain antibody fragments recognising VP2 of foot-and-mouth disease virus serotypes A, O, and SAT 2

Virology. 2019 Jul:533:145-154. doi: 10.1016/j.virol.2019.05.012. Epub 2019 May 25.

Abstract

Foot-and-mouth disease virus (FMDV) exhibits a high degree of antigenic diversity among its serotypes, requiring several anti-FMDV antibodies for its laboratory diagnosis, which complicated the used techniques. To conquer this cumbersome, we developed a new panel of different single-chain fragment variable (scFv) for serotype-independent detection of FMDV. The recombinant VP2 capsid protein, as a relatively conserved protein among FMDV serotypes, was expressed in E. Coli, and injected in mice. Spleen's RNA was extracted for isolating the coding sequences of IgG variable domains that were assembled into repertoires of scFv. Phage library displaying scFv was constructed with ∼1.9 × 108 plaque forming units. Characterization of the library showed eight of unique scFvs, which were expressed as bacterial periplasmic proteins with apparent molecular weight of ∼27 kDa. Our data revealed the broad-spectrum binding affinity of the eight scFvs as both coating and tracing antibodies to FMDV serotypes A, O, and SAT 2.

Keywords: Foot-and-mouth disease virus (FMDV); Phage-display; Serotype-independent; Single chain fragment variable (scFv); Splicing overlap extension (SOE)-PCR; VP2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Viral / analysis
  • Antibodies, Viral / immunology
  • Capsid Proteins / immunology*
  • Female
  • Foot-and-Mouth Disease / diagnosis
  • Foot-and-Mouth Disease / virology*
  • Foot-and-Mouth Disease Virus / classification
  • Foot-and-Mouth Disease Virus / genetics
  • Foot-and-Mouth Disease Virus / immunology
  • Foot-and-Mouth Disease Virus / isolation & purification*
  • Humans
  • Immunization
  • Mice
  • Mice, Inbred BALB C
  • Peptide Library
  • Serogroup
  • Single-Chain Antibodies / analysis*
  • Single-Chain Antibodies / genetics
  • Single-Chain Antibodies / immunology

Substances

  • Antibodies, Viral
  • Capsid Proteins
  • Peptide Library
  • Single-Chain Antibodies
  • VP2 protein, Foot-and-mouth disease virus