Phage Display and Selections on Cells

Methods Mol Biol. 2018:1701:321-330. doi: 10.1007/978-1-4939-7447-4_17.

Abstract

Antibody identification by phage display on protein or peptide targets is well established and many protocols are available. But there are many targets that cannot be expressed recombinantly or, like peptides, do not reflect correct folding of the protein. Most of these targets are cell surface receptors. Here, we describe a protocol for a panning strategy on cells to obtain specific binders to cell surface receptors. A depletion step is included to prevent enrichment of antibodies that bind to unwanted targets. Each step of the protocol is explained and variations of this protocol are given.

Keywords: Antibody engineering; Conformational epitope; Flow cytometry; Human antibodies; Phage display; Recombinant antibodies; Therapeutic antibodies; scFv.

MeSH terms

  • Cell Line
  • Cloning, Molecular / methods*
  • Gene Library*
  • Humans
  • Peptide Library*
  • Receptors, Cell Surface / chemistry*
  • Receptors, Cell Surface / immunology
  • Single-Chain Antibodies / chemistry*
  • Single-Chain Antibodies / genetics*
  • Single-Chain Antibodies / immunology

Substances

  • Peptide Library
  • Receptors, Cell Surface
  • Single-Chain Antibodies