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. 1994 May 1;13(9):2236–2243. doi: 10.1002/j.1460-2075.1994.tb06501.x

A general strategy to identify mimotopes of pathological antigens using only random peptide libraries and human sera.

A Folgori 1, R Tafi 1, A Meola 1, F Felici 1, G Galfré 1, R Cortese 1, P Monaci 1, A Nicosia 1
PMCID: PMC395079  PMID: 7514533

Abstract

A strategy to identify disease-specific epitopes from phage-displayed random peptide libraries using human sera is described. Peptides on phage (phagotopes) that react with antibodies present in patient sera are purified from > 10(7) different sequences by affinity selection and immunological screening of plaques. Disease-specific phagotopes can be identified out of this pool through an 'antigen independent' procedure which avails itself only of patient and normal human sera. Using this strategy, we have selected antigenic mimics (mimotopes) of two different epitopes from the human hepatitis B virus envelope protein (HBsAg). We could show that a humoral response to these mimotopes is widespread in the immunized population, suggesting that the strategy identifies phagotopes that have a potential role as diagnostic reagents. Immunization of mice with the selected phagotopes elicited a strong specific response against the HBsAg. These results open new inroads into disease-related epitope discovery and provide the potential for vaccine development without a requirement for the use of, or even information about, the aetiological agent or its antigens.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Blond-Elguindi S., Cwirla S. E., Dower W. J., Lipshutz R. J., Sprang S. R., Sambrook J. F., Gething M. J. Affinity panning of a library of peptides displayed on bacteriophages reveals the binding specificity of BiP. Cell. 1993 Nov 19;75(4):717–728. doi: 10.1016/0092-8674(93)90492-9. [DOI] [PubMed] [Google Scholar]
  2. Christian R. B., Zuckermann R. N., Kerr J. M., Wang L., Malcolm B. A. Simplified methods for construction, assessment and rapid screening of peptide libraries in bacteriophage. J Mol Biol. 1992 Oct 5;227(3):711–718. doi: 10.1016/0022-2836(92)90219-a. [DOI] [PubMed] [Google Scholar]
  3. Cohen I. R., Young D. B. Autoimmunity, microbial immunity and the immunological homunculus. Immunol Today. 1991 Apr;12(4):105–110. doi: 10.1016/0167-5699(91)90093-9. [DOI] [PubMed] [Google Scholar]
  4. Cwirla S. E., Peters E. A., Barrett R. W., Dower W. J. Peptides on phage: a vast library of peptides for identifying ligands. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6378–6382. doi: 10.1073/pnas.87.16.6378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Devlin J. J., Panganiban L. C., Devlin P. E. Random peptide libraries: a source of specific protein binding molecules. Science. 1990 Jul 27;249(4967):404–406. doi: 10.1126/science.2143033. [DOI] [PubMed] [Google Scholar]
  6. Felici F., Castagnoli L., Musacchio A., Jappelli R., Cesareni G. Selection of antibody ligands from a large library of oligopeptides expressed on a multivalent exposition vector. J Mol Biol. 1991 Nov 20;222(2):301–310. doi: 10.1016/0022-2836(91)90213-p. [DOI] [PubMed] [Google Scholar]
  7. Felici F., Luzzago A., Folgori A., Cortese R. Mimicking of discontinuous epitopes by phage-displayed peptides, II. Selection of clones recognized by a protective monoclonal antibody against the Bordetella pertussis toxin from phage peptide libraries. Gene. 1993 Jun 15;128(1):21–27. doi: 10.1016/0378-1119(93)90148-v. [DOI] [PubMed] [Google Scholar]
  8. Greenwood J., Willis A. E., Perham R. N. Multiple display of foreign peptides on a filamentous bacteriophage. Peptides from Plasmodium falciparum circumsporozoite protein as antigens. J Mol Biol. 1991 Aug 20;220(4):821–827. doi: 10.1016/0022-2836(91)90354-9. [DOI] [PubMed] [Google Scholar]
  9. Hammer J., Valsasnini P., Tolba K., Bolin D., Higelin J., Takacs B., Sinigaglia F. Promiscuous and allele-specific anchors in HLA-DR-binding peptides. Cell. 1993 Jul 16;74(1):197–203. doi: 10.1016/0092-8674(93)90306-b. [DOI] [PubMed] [Google Scholar]
  10. Keller P. M., Arnold B. A., Shaw A. R., Tolman R. L., Van Middlesworth F., Bondy S., Rusiecki V. K., Koenig S., Zolla-Pazner S., Conard P. Identification of HIV vaccine candidate peptides by screening random phage epitope libraries. Virology. 1993 Apr;193(2):709–716. doi: 10.1006/viro.1993.1179. [DOI] [PubMed] [Google Scholar]
  11. Levi S., Cesareni G., Arosio P., Lorenzetti R., Soria M., Sollazzo M., Albertini A., Cortese R. Characterization of human ferritin H chain synthetized in Escherichia coli. Gene. 1987;51(2-3):269–274. doi: 10.1016/0378-1119(87)90315-5. [DOI] [PubMed] [Google Scholar]
  12. Luzzago A., Felici F., Tramontano A., Pessi A., Cortese R. Mimicking of discontinuous epitopes by phage-displayed peptides, I. Epitope mapping of human H ferritin using a phage library of constrained peptides. Gene. 1993 Jun 15;128(1):51–57. doi: 10.1016/0378-1119(93)90152-s. [DOI] [PubMed] [Google Scholar]
  13. McLafferty M. A., Kent R. B., Ladner R. C., Markland W. M13 bacteriophage displaying disulfide-constrained microproteins. Gene. 1993 Jun 15;128(1):29–36. doi: 10.1016/0378-1119(93)90149-w. [DOI] [PubMed] [Google Scholar]
  14. Minenkova O. O., Ilyichev A. A., Kishchenko G. P., Petrenko V. A. Design of specific immunogens using filamentous phage as the carrier. Gene. 1993 Jun 15;128(1):85–88. doi: 10.1016/0378-1119(93)90157-x. [DOI] [PubMed] [Google Scholar]
  15. O'Neil K. T., Hoess R. H., Jackson S. A., Ramachandran N. S., Mousa S. A., DeGrado W. F. Identification of novel peptide antagonists for GPIIb/IIIa from a conformationally constrained phage peptide library. Proteins. 1992 Dec;14(4):509–515. doi: 10.1002/prot.340140411. [DOI] [PubMed] [Google Scholar]
  16. Oldenburg K. R., Loganathan D., Goldstein I. J., Schultz P. G., Gallop M. A. Peptide ligands for a sugar-binding protein isolated from a random peptide library. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5393–5397. doi: 10.1073/pnas.89.12.5393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Purcell R. H., Gerin J. L. Prospects for second and third generation hepatitis B vaccines. Hepatology. 1985 Jan-Feb;5(1):159–163. doi: 10.1002/hep.1840050130. [DOI] [PubMed] [Google Scholar]
  18. Scott J. K., Loganathan D., Easley R. B., Gong X., Goldstein I. J. A family of concanavalin A-binding peptides from a hexapeptide epitope library. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5398–5402. doi: 10.1073/pnas.89.12.5398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Scott J. K., Smith G. P. Searching for peptide ligands with an epitope library. Science. 1990 Jul 27;249(4967):386–390. doi: 10.1126/science.1696028. [DOI] [PubMed] [Google Scholar]
  20. Smith G. P. Surface presentation of protein epitopes using bacteriophage expression systems. Curr Opin Biotechnol. 1991 Oct;2(5):668–673. doi: 10.1016/0958-1669(91)90032-z. [DOI] [PubMed] [Google Scholar]
  21. Stephen C. W., Lane D. P. Mutant conformation of p53. Precise epitope mapping using a filamentous phage epitope library. J Mol Biol. 1992 Jun 5;225(3):577–583. doi: 10.1016/0022-2836(92)90386-x. [DOI] [PubMed] [Google Scholar]
  22. Tsunetsugu-Yokota Y., Tatsumi M., Robert V., Devaux C., Spire B., Chermann J. C., Hirsch I. Expression of an immunogenic region of HIV by a filamentous bacteriophage vector. Gene. 1991 Mar 15;99(2):261–265. doi: 10.1016/0378-1119(91)90136-y. [DOI] [PubMed] [Google Scholar]
  23. Willis A. E., Perham R. N., Wraith D. Immunological properties of foreign peptides in multiple display on a filamentous bacteriophage. Gene. 1993 Jun 15;128(1):79–83. doi: 10.1016/0378-1119(93)90156-w. [DOI] [PubMed] [Google Scholar]
  24. de la Cruz V. F., Lal A. A., McCutchan T. F. Immunogenicity and epitope mapping of foreign sequences via genetically engineered filamentous phage. J Biol Chem. 1988 Mar 25;263(9):4318–4322. [PubMed] [Google Scholar]

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