Affinity Selection Using Filamentous Phage Display

  • Mari Ann Kulseth
  • Annette Fagerlund
  • Astrid Hilde Myrset
Part of the Methods in Molecular Biology book series (MIMB, volume 1088)


Display of peptides on filamentous phage, phage display, is an in vitro selection technique well suited for identification of therapeutic peptide binders for a huge variety of protein targets. The peptides are identified in a process where phage libraries are subjected to affinity selection towards a particular protein target. A successful outcome of an affinity selection is dependent on proper surveillance of the phage life cycle, to make sure that the selection is based on affinity for the target, not on bias in phage propagation rate. In this chapter we present two approaches for protein target presentation and a protocol for phage rescue and propagation, which includes several controls to ensure that all phages initially eluted from the protein target are given equal conditions during the following amplification and selection steps.

Key words

Phage display Affinity selection Peptides VEGFR2 MPP9 



This work was performed at and supported by Medical Diagnostics Research, GE Healthcare.


  1. 1.
    Fagerlund A, Myrset AH, Kulseth MA (2008) Construction and characterization of a 9-mer phage display pVIII-library with regulated peptide density. Appl Microbiol Biotechnol 80:925–936PubMedCrossRefGoogle Scholar
  2. 2.
    Jacobson A (1972) Role of F pili in the penetration of bacteriophage fl. J Virol 10:835–843PubMedGoogle Scholar
  3. 3.
    Russel M, Lowmann HB, Clackson T (2004) Introduction to phage biology and phage display. In: Clackson T, Lowmann HB (eds) Phage display: a practical approach. Oxford University Press, New York, pp 1–26Google Scholar
  4. 4.
    Beekwilder J, Rakonjac J, Jongsma M, Bosch D (1999) A phagemid vector using the E. coli phage shock promoter facilitates phage display of toxic proteins. Gene 228:23–31PubMedCrossRefGoogle Scholar
  5. 5.
    Barrett RW, Cwirla SE, Ackerman MS, Olson AM, Peters EA, Dower WJ (1992) Selective enrichment and characterization of high affinity ligands from collections of random peptides on filamentous phage. Anal Biochem 204:357–364PubMedCrossRefGoogle Scholar
  6. 6.
    Sidhu SS, Lowman HB, Cunningham BC, Wells JA (2000) Phage display for selection of novel binding peptides. Methods Enzymol 328:333–358PubMedCrossRefGoogle Scholar
  7. 7.
    Kay BK, Kasanov J, Yamabhai M (2001) Screening phage-displayed combinatorial peptide libraries. Methods 24:240–246PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • Mari Ann Kulseth
    • 1
  • Annette Fagerlund
    • 2
  • Astrid Hilde Myrset
    • 3
  1. 1.Department of Medical Genetics, RikshospitaletOslo University HospitalOsloNorway
  2. 2.Department of Pharmaceutical BiosciencesUniversity of OsloOsloNorway
  3. 3.Omegatri AS, Oslo Innovation CentreOsloNorway

Personalised recommendations