A One-Step Process for the Construction of Phage Display scFv and VHH Libraries

Mol Biotechnol. 2020 Apr;62(4):228-239. doi: 10.1007/s12033-020-00236-0.

Abstract

In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes. We show that single chain variable fragment (scFv) libraries with adequate qualities can readily be cloned in a 'scar-less' manner and that the isolation of antigen-specific antibodies from immunized chickens is feasible within three selection rounds. Moreover, we demonstrate the general applicability of this method by rapidly constructing and panning VHH single domain antibody phage display libraries from immunized Llama repertoires.

Keywords: Antibody; Golden gate cloning; Phage display; Single domain antibody; Type IIs enzyme; VHH; scFv.

MeSH terms

  • Animals
  • Antibodies / immunology
  • Antibodies / isolation & purification
  • Bacteriophages / genetics
  • Camelids, New World
  • Cell Surface Display Techniques / methods*
  • Chickens
  • Deoxyribonucleases, Type II Site-Specific
  • ErbB Receptors / immunology
  • Escherichia coli
  • Single-Chain Antibodies / genetics*
  • Single-Chain Antibodies / immunology
  • Single-Domain Antibodies / genetics*
  • Single-Domain Antibodies / immunology

Substances

  • Antibodies
  • Single-Chain Antibodies
  • Single-Domain Antibodies
  • EGFR protein, human
  • ErbB Receptors
  • Deoxyribonucleases, Type II Site-Specific