Advertisement

Phage Display

  • Konstantin PetropoulosEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 901)

Abstract

Phage display has emerged as one of the leading technologies for the selection and generation of highly specific antibodies, offering a number of advantages over traditional ways of antibody generation such as mouse hybridoma techniques. While there are various possibilities to conduct phage display, selection of antibodies via solution panning is an elegant way to circumvent conformation changes of antigen, which may arise when performing panning with antigen immobilized on a solid surface. Here, a standard solution panning procedure using a Fab based antibody library including primary screening for selectivity is described.

Key words

Antibody fragment Phage display Antibody library Fab Solution panning 

References

  1. 1.
    Lindenmann J (1984) Senior overviews. Scand J Immunol 19:281–285PubMedCrossRefGoogle Scholar
  2. 2.
    Emil von Behring – Biography. Nobelprize.org. http://www.nobeprize.org/nobel_prizes/medicine/laureates/1901/behring.html. Accessed 16 Feb 2012
  3. 3.
    Kohler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495–497PubMedCrossRefGoogle Scholar
  4. 4.
    Bradbury AR, Sidhu S, Dubel S, McCafferty J (2011) Beyond natural antibodies: the power of in vitro display technologies. Nat Biotechnol 29:245–254PubMedCrossRefGoogle Scholar
  5. 5.
    Smith GP (1985) Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. Science 228:1315–1317PubMedCrossRefGoogle Scholar
  6. 6.
    Bradbury AR, Marks JD (2004) Antibodies from phage antibody libraries. J Immunol Methods 290:29–49PubMedCrossRefGoogle Scholar
  7. 7.
    Hoogenboom HR (2002) Overview of antibody phage-display technology and its applications. Methods Mol Biol 178:1–37PubMedGoogle Scholar
  8. 8.
    Knappik A, Ge L, Honegger A et al (2000) Fully synthetic human combinatorial antibody libraries (HuCAL) based on modular consensus frameworks and CDRs randomized with trinucleotides. J Mol Biol 296:57–86PubMedCrossRefGoogle Scholar
  9. 9.
    Van den Brulle J, Fischer M, Langmann T et al (2008) A novel solid phase technology for high-throughput gene synthesis. Biotechniques 45:340–343PubMedCrossRefGoogle Scholar
  10. 10.
    Burton DR, Scott JK, Silverman GJ (2001) Phage display. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USAGoogle Scholar
  11. 11.
    Finlay WJ, Bloom L, Cunningham O (2011) Optimized generation of high-affinity, high-specificity single-chain Fv antibodies from multiantigen immunized chickens. Methods Mol Biol 681:87–101PubMedCrossRefGoogle Scholar
  12. 12.
    Kotlan B, Glassy MC (2009) Antibody phage display: overview of a powerful technology that has quickly translated to the clinic. Methods Mol Biol 562:1–15PubMedCrossRefGoogle Scholar
  13. 13.
    Rothe C, Urlinger S, Lohning C et al (2008) The human combinatorial antibody library HuCAL GOLD combines diversification of all six CDRs according to the natural immune system with a novel display method for efficient selection of high-affinity antibodies. J Mol Biol 376:1182–1200PubMedCrossRefGoogle Scholar
  14. 14.
    Løset GÅ, Bogen B, Sandlie I (2011) Expanding the versatility of phage display I: efficient display of peptide-tags on protein VII of the filamentous phage. PLoS One 6(2):e14702PubMedCrossRefGoogle Scholar
  15. 15.
    Barat B, Wu AM (2007) Metabolic biotinylation of recombinant antibody by biotin ligase retained in the endoplasmic reticulum. Biomol Eng 24:283–291PubMedCrossRefGoogle Scholar
  16. 16.
    Thermo Scientific Avidin-Biotin Technical Handbook (2009) http://www.piercenet.com/browse.cfm?fldID=84EBE112-F871-4CA5-807F-47327153CFCB
  17. 17.
    Hermanson GT (2008) Bioconjugate techniques, 2nd edn. Academic, New York, NYGoogle Scholar
  18. 18.
    Clackson T, Lowman HB (2004) Phage display: a practical approach. Oxford University Press, OxfordGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.MorphoSys AGMartinsried/PlaneggGermany

Personalised recommendations