Skip to main content
SpringerLink
Log in

A combinatorial method for constructing libraries of long peptides displayed by filamentous phage

  • Research Papers
  • Published:
Molecular Diversity Aims and scope Submit manuscript

Summary

We describe the construction and screening of a random peptide library displayed by filamentous phage. The peptides are expressed in multiple copies on the filamentous phage M13 as amino-terminal fusions with the major coat protein, the product of gene VIII. These libraries are efficiently screened for reactive peptides, using a combination of panning in solution followed by a plaque lift assay. Advantages of this system are that both high- and low-affinity phage clones are simultaneously identified and the analysis of non-reactive phage is minimized. The vector system utilized to construct this library enables it to be used for the construction of peptide libraries employing a combinatorial cloning strategy. This feature makes it especially suitable for construction of peptide libraries using codon-based oligonucleotide synthesis. The vectors also allow rapid optimization and modification of lead peptides by codon-based mutagenesis. A 20-amino acid long random peptide library of 1 × 109 members was constructed and screened for peptides that bound to (i) a monoclonal antibody recognizing the amino-terminus of β-endorphin; (ii) a monoclonal antibody recognizing a peptide epitope derived from the v -ros oncogene product; and (iii) the constant region of murine IgG2b. The approach described here provides a means for the construction of customized libraries that can be screened with a variety of target molecules.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Brenner, S. and Lerner, R.A.,Encoded combinatorial chemistry, Proc. Natl. Acad. Sci. USA, 89 (1992) 5381–5383.

    Google Scholar 

  2. Kenan, D.J., Tsai, D.E. and Keene, J.D.,Exploring molecular diversity with combinatorial shape libraries, Trends Biochem. Sci., 19 (1994) 57–64.

    Google Scholar 

  3. Scott, J.K.,Discovering peptide ligands using epitope libraries, Trends Biochem. Sci., 17 (1992) 241–245.

    Google Scholar 

  4. Scott, J.K. and Craig, L.,Random peptide libraries, Curr. Opin. Biotechnol., 5 (1994) 40–48.

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  8. Fodor, S.P.A., Read, J.L., Pirrung, M.C., Stryer, L., Lu, A.T. and Solas, D.,Light-directed, spatially addressable parallel chemical synthesis, Science, 251 (1991) 767–773.

    Google Scholar 

  9. Houghten, R.A., Pinilla, C., Blondelle, S.E., Appel, J.R., Dooley, C.T. and Cuervo, J.H.,Generation and use of synthetic peptide combinatorial libraries for basic research and drug discovery, Nature, 354 (1991) 84–86.

    Google Scholar 

  10. Lam, K.S., Salmon, S.E., Hersh, E.M., Hruby, V.J., Kazmierski, W.M. and Knapp, R.J.,A new type of synthetic peptide library for identifying ligand-binding activity, Nature, 354 (1991) 82–84.

    Google Scholar 

  11. Jayawickreme, C.K., Graminski, G.F., Quillan, J.M. and Lerner, M.,Creation and functional screening of a multi-use peptide library, Proc. Natl. Acad. Sci. USA, 91 (1994) 1614–1618.

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  14. De la Cruz, V.F., Lal, A. and McCutchan, T.F.,Immunogenicity and epitope mapping of foreign sequences via genetically engineered filamentous phage, J. Biol. Chem., 263 (1988) 4318–4322.

    Google Scholar 

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

    Google Scholar 

  16. Model, P. and Russel, M.,Filamentous Bacteriophage, In Calendar, R. (Ed.) The Bacteriophages, Vol. 2, Plenum Press, New York, NY, 1988, pp. 375–456.

    Google Scholar 

  17. Felici, F., Castagnoli, L., Musacchio, A., Jappelli, R. and Cesareni, G.,Selection of antibody ligands from a large library of oligopeptides expressed on a multivalent exposition vector, J. Mol. Biol., 222 (1991) 301–310.

    Google Scholar 

  18. Greenwood, J., Willis, A.E. and Perham, R.N.,Multiple display of foreign peptides on a filamentous bacteriophage. Peptides from Plasmodium falciparum circumsporozoite protein as antigens, J. Mol. Biol., 220 (1991) 821–827.

    Google Scholar 

  19. Kay, B.K., Adey, N.B., He, Y.-S., Manfredi, J.P., Mataragnon, A.H. and Fowlkes, D.M.,An M13 phage library displaying random 38-amino-acid peptides as a source of novel sequences with affinity to selected targets, Gene, 128 (1993) 59–65.

    Google Scholar 

  20. Huse, W.D., Sastry, L., Iverson, S.A., Kang, A.S., Alting-Mees, M., Burton, D.R., Bencovic, S.J. and Lerner, R.A.,Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda, Science, 246 (1989) 1275–1281.

    Google Scholar 

  21. McCafferty, I., Griffiths, A.D., Winter, G. and Chiswell, D.J.,Phage antibodies: Filamentous phage displaying antibody variable domains, Nature, 348 (1990) 552–554.

    Google Scholar 

  22. JOhnson, K.S. and Chiswell, D.J.,Human antibody engineering, Curr. Opin. Struct. Biol., 3 (1993) 564–571.

    Google Scholar 

  23. Barbas, C.F.,Recent advances in phage display, Curr. Opin. Struct. Biol., 4 (1993) 526–530.

    Google Scholar 

  24. Hoess, R.H.,Phage display of peptides and protein domains, Curr. Opin. Struct. Biol., 3 (1993) 572–579.

    Google Scholar 

  25. Makowski, L.,Phage display: Structure, assembly and engineering of filamentous bacteriophage M13, Curr. Opin. Struct. Biol., 4 (1994) 225–230.

    Google Scholar 

  26. Maruyama, I.N., Maruyama, H.I. and Brenner, S.,λfoo: A λ phage vector for the expression of foreign proteins, Proc. Natl. Acad. Sci. USA, 91 (1994) 8273–8277.

    Google Scholar 

  27. Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.G. and Struhl, K. (Eds.) Current Protocols in Molecular Biology, Wiley, New York, NY, 1987.

    Google Scholar 

  28. Kunkel, T.A., Roberts, J. and Zakour, R.,Rapid and efficient sitespecific mutagenesis without phenotypic selection, Methods Enzymol., 154 (1987) 367–382.

    Google Scholar 

  29. Yanisch-Perron, C., Vieira, J. and Messing, J.,Improved M13 phage cloning vectors and host strains: Nucleotide sequences of the M13, MP18 and PUC19 vectors, Gene, 33 (1985) 103–119.

    Google Scholar 

  30. Inouye, H., Barnes, W. and Beckwith, J.,Signal sequence of alkaline phosphatase of Escherichia coli, J. Bacteriol., 149 (1982) 434–439.

    Google Scholar 

  31. Folgori, A., Tafi, R., Meola, A., Felici, F., Galfre', G., Cortese, R., Monaci, P. and Nicosia, A.,A general strategy to identify mimotopes of pathological antigens using only random peptide libraries and human sera, EMBO J., 13 (1994) 2236–2243.

    Google Scholar 

  32. Tabor, S. and Richardson, C.C.,DNA sequence analysis with a modified bacteriophage T7 DNA polymerase, Proc. Natl. Acad. Sci. USA, 84 (1987) 4767–4771.

    Google Scholar 

  33. Huse, W.D., Stinchcombe, T.J., Glaser, S.M., Starr, L., MacLean, M., Hellstrom, K.E., Hellstrom, I. and Yelton, D.E.,Application of a filamentous phage pVIII fusion protein system suitable for efficient production, screening, and mutagenesis of F(ab) antibody fragments, J. Immunol., 149 (1992) 3914–3920.

    Google Scholar 

  34. Gramsch, C., Meo, T., Riethmuller, G. and Herz, A.,Binding characteristics of a monoclonal β-endorphin antibody recognizing the N-terminus of opioid peptides, J. Neurochem., 40 (1983) 1220–1226.

    Google Scholar 

  35. Schägger, H. and von Jagow, G.,Tricine-sodium dodecyl sulfatepolyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa, Anal. Biochem., 166 (1987) 368–379.

    Google Scholar 

  36. Watson, J.D., Hopkins, N.H., Roberts, J.W., Steitz, J.A. and Weiner, A.M., Molecular Biology of the Gene, Benjamin/Cummings, Menlo Park, CA, 1988, p. 370.

    Google Scholar 

  37. Taylor, W.R.,The classification of amino acid conservation, J. Theor. Biol., 119 (1986) 205–218.

    Google Scholar 

  38. Walz, G., Aruffo, A., Kolanus, W., Bevilacqua, M. and Seed, B.,Recognition by ELAM-1 of the sialyl-Le X determinant on myeloid and tumor cells, Science, 250 (1990) 1132–1135.

    Google Scholar 

  39. Glaser, S.M., Yelton, D.E. and Huse, W.D.,Antibody engineering by codon-based mutagenesis in a filamentous phage vector system, J. Immunol., 149 (1992) 3903–3913.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Ixsys Inc., 3550 Dunhill Street, 92121, San Diego, CA, USA

    Tapio Haaparanta & William D. Huse

Authors
  1. Tapio Haaparanta
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. William D. Huse
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Haaparanta, T., Huse, W.D. A combinatorial method for constructing libraries of long peptides displayed by filamentous phage. Mol Divers 1, 39–52 (1995). https://doi.org/10.1007/BF01715808

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01715808

Keywords

Search

Navigation