Protein Arrays

Volume 264 of the series Methods in Molecular Biology pp 47-60

Target-Assisted Iterative Screening of Phage Surface Display cDNA Libraries

  • Alexei KurakinAffiliated withBuck Institute for Age Research
  • , Susan WuAffiliated withBuck Institute for Age Research
  • , Dale E. BredesenAffiliated withBuck Institute for Age Research

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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.

Key Words

TAIS phage surface display cDNA library screening format molecular repertoires protein-protein interactions