Antibody Phage Display: Antibody Selection in Solution Using Biotinylated Antigens

Methods Mol Biol. 2020:2070:143-155. doi: 10.1007/978-1-4939-9853-1_8.

Abstract

Antibody phage display is the most used in vitro technology to generate recombinant, mainly human, antibodies as tools for research, for diagnostic assays, and for therapeutics. Up to now (autumn 2018), eleven FDA/EMA-approved therapeutic antibodies were developed using phage display, including the world best-selling antibody adalimumab.A key to generate successfully human antibodies in vitro is the choice of the most appropriate antibody selection method, for our goal. In this book chapter, we describe the antibody selection process (panning) in solution and its advantages over panning on immobilized antigens. Detailed protocols on the panning procedure and the screening of monoclonal binders are given.

Keywords: Antibody; Panning; Phage display; ScFv.

MeSH terms

  • Antigens / chemistry*
  • Biotin / chemistry*
  • Biotinylation
  • Humans
  • Peptide Library*
  • Single-Chain Antibodies* / chemistry
  • Single-Chain Antibodies* / genetics

Substances

  • Antigens
  • Peptide Library
  • Single-Chain Antibodies
  • Biotin