Phage selection of bicyclic peptides

Methods. 2013 Mar 15;60(1):46-54. doi: 10.1016/j.ymeth.2012.12.008. Epub 2013 Jan 10.


Bicyclic peptides are small, constrained peptides that can bind with high affinity and selectivity to protein targets. Their small size provides a number of advantages over larger protein-based ligands, including access to chemical synthesis, better tissue penetration, and a wider choice of application routes. Bicyclic peptide ligands can be identified using phage display technology with moderate effort and cost. Here we provide step-by-step protocols for the isolation of bicyclic peptide ligands using phage display. These protocols have been successfully used in our laboratory for the generation of high-affinity binders to a variety of protein targets. We describe library generation, affinity selection and ligand characterization, and provide troubleshooting advice concerning frequent problems.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Bacteriophages*
  • Cell Surface Display Techniques*
  • Ligands
  • Peptide Library*
  • Peptides, Cyclic*


  • Ligands
  • Peptide Library
  • Peptides, Cyclic