Construction of a novel phage display antibody library against Fasciola hepatica, and generation of a single-chain variable fragment specific for F. hepatica cathepsin L1

Exp Parasitol. 2019 Mar:198:87-94. doi: 10.1016/j.exppara.2019.02.001. Epub 2019 Feb 11.

Abstract

Phage display technology to produce recombinant monoclonal antibodies or antibody fragments permits the identification of sought after antibodies in short time frames at low cost along with direct and rapid selection for antibody characteristics. Monoclonal antibodies can facilitate the identification and characterisation of parasite molecules that function at the host-parasite interface to help understand at the molecular level the biology of the parasite and disease progression, which often leads to new drug targets, diagnostic antigens or vaccine candidates. The trematode Fasciola hepatica is an important veterinary and human parasite. In this work, we infected rats with F. hepatica and amplified the generated antibody repertoire to produce a single-chain variable fragment (scFv) phage display library. The library was used to identify a scFv that recognises cathepsin L1, a major component of the adult parasites excretory/secretory material and an important vaccine candidate. This is the first report of the construction of a phage display antibody library from a F. hepatica infected host, and also the first instance such a library has been used to identify an affinity-matured monoclonal antibody fragment that binds to a F. hepatica antigen. The scFv library and methods detailed should facilitate future research characterising F. hepatica antigens.

Keywords: Cathepsin; Fasciola hepatica; Phage display; Single-chain variable fragment (scFv).

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Helminth / immunology*
  • Antibodies, Monoclonal / immunology
  • Antigens, Helminth / immunology
  • Cathepsins / immunology*
  • Cell Surface Display Techniques
  • Cloning, Molecular
  • Electrophoresis, Agar Gel
  • Fasciola hepatica / immunology*
  • Gene Library
  • Immunoblotting
  • Immunoglobulin Variable Region / chemistry
  • Peptide Library*
  • Polymerase Chain Reaction
  • Rats
  • Rats, Inbred Lew
  • Single-Chain Antibodies / genetics
  • Single-Chain Antibodies / immunology

Substances

  • Antibodies, Helminth
  • Antibodies, Monoclonal
  • Antigens, Helminth
  • Immunoglobulin Variable Region
  • Peptide Library
  • Single-Chain Antibodies
  • Cathepsins
  • cathepsin L1, Fasciola hepatica