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Epitope Mapping Protocols pp 181–201Cite as

Epitope Mapping Using Phage Display Peptide Libraries

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 524))

Summary

Phage libraries displaying millions of peptides with randomized sequences are extremely useful tools for mapping antibody epitopes. In many cases, antibodies are able to select peptides with reasonable affinity for their combining sites (paratopes) from these libraries. Ideally, consensus motives can be deduced from multiple peptide sequences and matched to areas of the antigen against which the antibody was raised. That way, critical components of the antibody epitope can be defined. This chapter focuses on technical details of epitope mapping employing pre-made filamentous phage peptide display libraries. Examples are given for illustration.

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References

  1. Parmley, S. F. and Smith, G. P. (1988) Antibody-selectable filamentous fd phage vectors: affinity purification of target genes. Gene 73, 305–318.

    Article  PubMed  CAS  Google Scholar 

  2. Smith, G. P. (1985) Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. Science 228, 1315–1317.

    Article  PubMed  CAS  Google Scholar 

  3. Cwirla, S. E., Peters, E. A., Barrett, R. W., and Dower, W. J. (1990) Peptides on phage: a vast library of peptides for identifying ligands. Proc. Natl. Acad. Sci. USA 87, 6378–6382.

    Article  PubMed  CAS  Google Scholar 

  4. Devlin, J. J., Panganiban, L. C., and Devlin, P. E. (1990) Random peptide libraries: a source of specific protein binding molecules. Science 249, 404–406.

    Article  PubMed  CAS  Google Scholar 

  5. Scott, J. K. and Smith, G. P. (1990) Searching for peptide ligands with an epitope library. Science 249, 386–390.

    Article  PubMed  CAS  Google Scholar 

  6. Kehoe, J. W. and Kay, B. K. (2005) Filamentous phage display in the new millennium. Chem. Rev. 105, 4056–4072.

    Article  PubMed  CAS  Google Scholar 

  7. Smith, G. P. and Petrenko, V. A. (1997) Phage Display. Chem. Rev. 97, 391–410.

    Article  PubMed  CAS  Google Scholar 

  8. Pini, A., Giuliani, A., Ricci, C., Runci, Y., and Bracci, L. (2004) Strategies for the construction and use of peptide and antibody libraries displayed on phages. Curr. Protein Pept. Sci. 5, 487–496.

    Article  PubMed  CAS  Google Scholar 

  9. Rowley, M. J., O’Connor, K., and Wijeyewickrema, L. (2004) Phage display for epitope determination: a paradigm for identifying receptor-ligand interactions. Biotechnol. Annu. Rev. 10, 151–188.

    Article  PubMed  CAS  Google Scholar 

  10. Szardenings, M. (2003) Phage display of random peptide libraries: applications, limits, and potential. J. Recept. Signal Transduct. Res. 23, 307–349.

    Article  PubMed  CAS  Google Scholar 

  11. Sternberg, N. and Hoess, R. H. (1995) Display of peptides and proteins on the surface of bacteriophage lambda. Proc. Natl. Acad. Sci. USA 92, 1609–1613.

    Article  PubMed  CAS  Google Scholar 

  12. Efimov, V. P., Nepluev, I. V., and Mesyanzhinov, V. V. (1995) Bacteriophage T4 as a surface display vector. Virus Genes 10, 173–177.

    Article  PubMed  CAS  Google Scholar 

  13. Krumpe, L. R., Atkinson, A. J., Smythers, G. W., Kandel, A., Schumacher, K. M., McMahon, J. B., Makowski, L., and Mori, T. (2006) T7 lytic phage-displayed peptide libraries exhibit less sequence bias than M13 filamentous phage-displayed peptide libraries. Proteomics 6, 4210–4222.

    Article  PubMed  CAS  Google Scholar 

  14. Lindqvist, B. H. and Naderi, S. (1995) Peptide presentation by bacteriophage P4. FEMS Microbiol. Rev. 17, 33–39.

    Article  PubMed  CAS  Google Scholar 

  15. Stahl, S. and Uhlen, M. (1997) Bacterial surface display: trends and progress. Trends Biotechnol. 15, 185–192.

    Article  PubMed  CAS  Google Scholar 

  16. Smith, G. P. and Scott, J. K. (1993) Libraries of peptides and proteins displayed on filamentous phage. Methods Enzymol. 217, 228–257.

    Article  PubMed  CAS  Google Scholar 

  17. Bottger, V. (2001) Epitope mapping with random peptide libraries, in Antibody engineering (Kontermann, R. and Dubel, S., eds.), Springer Verlag, Heidelberg, Germany, pp. 460–472.

    Google Scholar 

  18. Bottger, V., Bottger, A., Lane, E. B., and Spruce, B. A. (1995) Comprehensive epitope analysis of monoclonal anti-proenkephalin antibodies using phage display libraries and synthetic peptides: revelation of antibody fine specificities caused by somatic mutations in the variable region genes. J. Mol. Biol. 247, 932–946.

    Article  PubMed  CAS  Google Scholar 

  19. Bottger, V. and Lane, E. B. (1994) A monoclonal antibody epitope on keratin 8 identified using a phage peptide library. J. Mol. Biol. 235, 61–67.

    Article  PubMed  CAS  Google Scholar 

  20. Bottger, V., Stasiak, P. C., Harrison, D. L., Mellerick, D. M., and Lane, E. B. (1995) Epitope mapping of monoclonal antibodies to keratin 19 using keratin fragments, synthetic peptides and phage peptide libraries. Eur. J. Biochem. 231, 475–485.

    Article  PubMed  CAS  Google Scholar 

  21. Kirschenhofer, A., Magdolen, V., Schmitt, M., Albrecht, S., Krol, J., Farthmann, J., Kopitz, C., Prezas, P., Kruger, A., Luther, T., and Bottger, V. (2003) Recombinant single chain antibody scFv-IIIF10 directed to human urokinase receptor. Recent Res. Devel. Cancer 5, 9–25.

    CAS  Google Scholar 

  22. D’Mello, F. and Howard, C. R. (2001) An improved selection procedure for the screening of phage display peptide libraries. J. Immunol. Methods 247, 191–203.

    Article  PubMed  Google Scholar 

  23. Bublil, E. M., Freund, N. T., Mayrose, I., Penn, O., Roitburd-Berman, A., Rubinstein, N. D., Pupko, T., and Gershoni, J. M. (2007) Stepwise prediction of conformational discontinuous B-cell epitopes using the Mapitope algorithm. Proteins 68, 294–304.

    Article  PubMed  CAS  Google Scholar 

  24. Gershoni, J. M., Roitburd-Berman, A., Siman-Tov, D. D., Tarnovitski Freund, N., and Weiss, Y. (2007) Epitope mapping: the first step in developing epitope-based vaccines. Biodrugs 21, 145–156.

    Article  PubMed  CAS  Google Scholar 

  25. Irving, M. B., Pan, O., and Scott, J. K. (2001) Random-peptide libraries and antigen-fragment libraries for epitope mapping and the development of vaccines and diagnostics. Curr. Opin. Chem. Biol. 5, 314–324.

    Article  PubMed  CAS  Google Scholar 

  26. Luther, T., Magdolen, V., Albrecht, S., Kasper, M., Riemer, C., Kessler, H., Graeff, H., Muller, M., and Schmitt, M. (1997) Epitope-mapped monoclonal antibodies as tools for functional and morphological analyses of the human urokinase receptor in tumor tissue. Am. J. Pathol. 150, 1231–1244.

    PubMed  CAS  Google Scholar 

  27. Spruce, B. A., Curtis, R., Wilkin, G. P., and Glover, D. M. (1990) A neuropeptide precursor in cerebellum: proenkephalin exists in subpopulations of both neurons and astrocytes. EMBO J. 9, 1787–1795.

    PubMed  CAS  Google Scholar 

  28. Kay, B. K., Kasanov, J., and Yamabhai, M. (2001) Screening phage-displayed combinatorial peptide libraries. Methods 24, 240–246.

    Article  PubMed  CAS  Google Scholar 

  29. Noren, K. A. and Noren, C. J. (2001) Construction of high-complexity combinatorial phage display peptide libraries. Methods 23, 169–178.

    Article  PubMed  CAS  Google Scholar 

  30. Menendez, A. and Scott, J. K. (2005) The nature of target-unrelated peptides recovered in the screening of phage-displayed random peptide libraries with antibodies. Anal. Biochem. 336, 145–157.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Wilex AG, Grillparzerstr. 10, 81675, Munich, Germany

    Volker Böttger

  2. Department Biology II, Ludwig-Maximilians-University Munich, Planegg-Martinsried, Germany

    Angelika Böttger

Authors
  1. Volker Böttger
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  2. Angelika Böttger
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Correspondence to Volker Böttger .

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    © 2009 Humana Press, a part of Springer Science+Business Media, LLC

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    Böttger, V., Böttger, A. (2009). Epitope Mapping Using Phage Display Peptide Libraries. In: Schutkowski, M., Reineke, U. (eds) Epitope Mapping Protocols. Methods in Molecular Biology™, vol 524. Humana Press. https://doi.org/10.1007/978-1-59745-450-6_13

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    • DOI: https://doi.org/10.1007/978-1-59745-450-6_13

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    • Publisher Name: Humana Press

    • Print ISBN: 978-1-934115-17-6

    • Online ISBN: 978-1-59745-450-6

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