ORF phage display to identify cellular proteins with different functions

Methods. 2012 Sep;58(1):2-9. doi: 10.1016/j.ymeth.2012.07.013. Epub 2012 Jul 23.


Open reading frame (ORF) phage display is a new branch of phage display aimed at improving its efficiency to identify cellular proteins with specific binding or functional activities. Despite the success of phage display with antibody libraries and random peptide libraries, phage display with cDNA libraries of cellular proteins identifies a high percentage of non-ORF clones encoding unnatural short peptides with minimal biological implications. This is mainly because of the uncontrollable reading frames of cellular proteins in conventional cDNA libraries. ORF phage display solves this problem by eliminating non-ORF clones to generate ORF cDNA libraries. Here I summarize the procedures of ORF phage display, discuss the factors influencing its efficiency, present examples of its versatile applications, and highlight evidence of its capability of identifying biologically relevant cellular proteins. ORF phage display coupled with different selection strategies is capable of delineating diverse functions of cellular proteins with unique advantages.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autoantigens / genetics
  • Bacteriophages / genetics
  • Cell Surface Display Techniques*
  • Cells, Cultured
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Gene Library
  • Genetic Vectors
  • Humans
  • Open Reading Frames*
  • Peptide Hydrolases / chemistry
  • Peptide Library*
  • Phagocytosis
  • Protein Interaction Mapping
  • Proteolysis
  • Proteome / genetics


  • Autoantigens
  • DNA, Complementary
  • Peptide Library
  • Proteome
  • Peptide Hydrolases