Pyrosequencing® of Phage Display Libraries for the Identification of Cell-Specific Targeting Ligands

  • Ahad A. Rahim
Part of the Methods in Molecular Biology™ book series (MIMB, volume 373)


High combinatorial phage display libraries have become an important tool in the search for ligand-receptor interactions. The advantage this approach offers is the ability to screen large repertoires of peptides, displayed on the coat proteins of bacteriophages, against a target at the same time. In addition, no prior knowledge is required of the target or the ligand. However, to characterize the peptides of interest a short length of the bacteriophage genome that encodes the peptide sequence requires DNA sequencing. The number of candidate bacteriophages can be large and so sequencing is expensive, timeconsuming, and laborious. Therefore, a methodology using Pyrosequencing® has been developed where 96-phage displaying a seven amino acid peptide can be analyzed simultaneously within 45 min and at a fraction of the cost associated with traditional automated Sanger sequencing.


  1. 1.
    Smith, G. P. (1985) Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. Science 228, 1315–1317.CrossRefPubMedGoogle Scholar
  2. 2.
    Fairbrother, W. J., Christinger, H. W., Cochran, A. G., et al. (1998) Novel peptides selected to bind vascular endothelial growth factor target the receptor-binding site. Biochemistry 37, 17,754–17,764.CrossRefPubMedGoogle Scholar
  3. 3.
    Healy, J. M., Murayama, O., Maeda, T., Yoshino, K., Sekiguchi, K., and Kikuchi, M. (1996) Peptide ligands for integrin alpha v beta 3 selected from random phage display libraries. Biochemistry 34, 3948–3955.CrossRefGoogle Scholar
  4. 4.
    Jost, P. J., Harbottle, R. P., Knight, A., Miller, A. D., Coutelle, C., and Schneider, H. (2001) A novel peptide, THALWHT, for the targeting of human airway epithelia. FEBS Lett. 489, 263–269.CrossRefPubMedGoogle Scholar
  5. 5.
    Nicklin, S. A., White, S. J., Watkins, S. J., Hawkins, R. E., and Baker, A. H. (2000) Selective targeting of gene transfer to vascular endothelial cells by use of peptides isolated by phage display. Circulation 102, 231–237.PubMedGoogle Scholar
  6. 6.
    Rahim, A., Coutelle,, C. and Harbottle, R. (2003) High-throughput Pyrosequencing of a phage display library for the identification of enriched target-specific peptides. Biotechniques 35, 317–324.PubMedGoogle Scholar
  7. 7.
    Pasqualini, R. and Ruoslahti, E. (1996) Organ targeting in vivo using phage display peptide libraries. Nature 380, 364–366.CrossRefPubMedGoogle Scholar
  8. 8.
    Arap, W., Kolonin, M. G., Trepel, M., et al. (2002) Steps toward mapping the human vasculature by phage display. Nat. Med. 8, 121–127.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  • Ahad A. Rahim
    • 1
  1. 1.Gene Therapy Group, Chester Beatty LaboratoriesInstitute of Cancer ResearchLondonUK

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