Affinity selection using filamentous phage display

Methods Mol Biol. 2014:1088:67-80. doi: 10.1007/978-1-62703-673-3_5.

Abstract

Display of peptides on filamentous phage, phage display, is an in vitro selection technique well suited for identification of therapeutic peptide binders for a huge variety of protein targets. The peptides are identified in a process where phage libraries are subjected to affinity selection towards a particular protein target. A successful outcome of an affinity selection is dependent on proper surveillance of the phage life cycle, to make sure that the selection is based on affinity for the target, not on bias in phage propagation rate. In this chapter we present two approaches for protein target presentation and a protocol for phage rescue and propagation, which includes several controls to ensure that all phages initially eluted from the protein target are given equal conditions during the following amplification and selection steps.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Chromatography, Affinity / methods*
  • Immobilized Proteins / metabolism
  • Inovirus / metabolism*
  • Molecular Sequence Data
  • Peptide Library*
  • Peptides / chemistry
  • Sequence Analysis, Protein
  • Solutions

Substances

  • Immobilized Proteins
  • Peptide Library
  • Peptides
  • Solutions