Rapid selection of high-affinity binders using ribosome display

Methods Mol Biol. 2012;805:261-86. doi: 10.1007/978-1-61779-379-0_15.

Abstract

Ribosome display has proven to be a powerful in vitro selection and evolution method for generating high-affinity binders from libraries of folded proteins. It has been successfully applied to single-chain Fv fragments of antibodies and alternative scaffolds, such as Designed Ankyrin Repeat Proteins (DARPins). High-affinity binders with new target specificity can be obtained from highly diverse DARPin libraries in only a few selection rounds. In this protocol, the selection from the library and the process of affinity maturation and off-rate selection are explained in detail.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Peptide Library
  • Polymerase Chain Reaction
  • Protein Binding
  • Protein Folding
  • Proteins / chemistry
  • Proteins / genetics*
  • Proteins / metabolism*
  • Ribosomes / genetics*
  • Ribosomes / metabolism*
  • Substrate Specificity

Substances

  • Peptide Library
  • Proteins