Target-assisted iterative screening of phage surface display cDNA libraries

Methods Mol Biol. 2004;264:47-60. doi: 10.1385/1-59259-759-9:047.

Abstract

The novel screening format, target-assisted iterative screening (TAIS), comprises a simple and rapid two-step procedure for in vitro affinity selection of specific binders from enormous molecular diversities to the target molecule of interest. This detailed protocol describes the application of TAIS to a T7 phage-displayed complementary DNA (cDNA) library with a protein domain as a target. Protocols for purification of the target as glutathione-S-transferase (GST) fusion protein and modifications of the purified target that are required for the screening complement the TAIS protocol. The described application is a method of choice for the researchers interested in the identification and characterization of novel protein-protein interactions mediated by peptide recognition domains.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Gene Library*
  • Humans
  • Peptide Library*
  • Protein Binding
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Peptide Library
  • Recombinant Fusion Proteins