1887

Abstract

This study describes the identification of the epitope recognized by the tobacco mosaic virus (TMV) coat protein (CP)-specific monoclonal antibody 29 (MAb29) by displaying a CP gene-fragment library on pVIII of filamentous phage M13. More than 80% of the clones isolated after one round of panning bound specifically to MAb29. DNA sequencing of ten randomly chosen MAb29-specific clones and subsequent sequence comparison revealed a common seven amino acid epitope (ELIRGTG) representing amino acids 131–137 of the TMV CP. The reactivity of MAb29 in competition ELISA towards glutathione -transferase fused to this epitope was stronger than that towards full-length wild-type TMV CP, confirming the epitope sequence determined by gene-fragment phage display. This demonstrated that gene-fragment libraries displayed on the phage surface as fusion proteins with the filamentous bacteriophage gene VIII are useful tools for rapid identification of linear epitopes recognized by MAbs.

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2001-01-01
2019-10-21
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References

  1. Al Moudallal, Z., Briand, J. P. & Van Regenmortel, M. H. V. ( 1982; ). Monoclonal antibodies as probes of the antigenic structure of tobacco mosaic virus. EMBO Journal 1, 1005-1010.
    [Google Scholar]
  2. Altschuh, D., Lesk, A. M., Bloomer, A. C. & Klug, A. ( 1987; ). Correlation of co-ordinated amino acid substitutions with function in viruses related to tobacco mosaic virus. Journal of Molecular Biology 193, 693-707.[CrossRef]
    [Google Scholar]
  3. Anderson, S. ( 1981; ). Shotgun DNA sequencing using cloned DNase I-generated fragments. Nucleic Acids Research 9, 3015-3027.[CrossRef]
    [Google Scholar]
  4. Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. & Struhl, K. (2000). Current Protocols in Molecular Biology. New York: John Wiley.
  5. Bloomer, A. C. & Butler, P. J. G. (1986). Tobacco mosaic virus – structure and self-assembly. In The Plant Viruses, vol. 2, pp. 4–57. New York: Plenum.
  6. Böttger, V., Stasiak, P. C., Harrison, D. L., Mellerick, D. M. & Lane, E. B. ( 1995; ). Epitope mapping of monoclonal antibodies to keratin 19 using keratin fragments, synthetic peptides and phage peptide libraries. European Journal of Biochemistry 231, 475-485.[CrossRef]
    [Google Scholar]
  7. Burritt, J. B., Bond, C. W., Doss, K. W. & Jesaitis, A. J. ( 1996; ). Filamentous phage display of oligopeptide libraries. Analytical Biochemistry 238, 1-13.[CrossRef]
    [Google Scholar]
  8. Cesareni, G. ( 1992; ). Peptide display on filamentous phage capsids. A new powerful tool to study protein–ligand interaction. FEBS Letters 307, 66-70.[CrossRef]
    [Google Scholar]
  9. Coligan, J. E., Kruisbeek, A. M., Margulies, D. H., Shevach, E. M. & Strober, W. (2000). Current Protocols in Immunology. New York: John Wiley.
  10. Cortese, R., Monaci, P., Nicosia, A., Luzzago, A., Felici, F., Galfre, G., Pessi, A., Tramontano, A. & Sollazzo, M. ( 1995; ). Identification of biologically active peptides using random libraries displayed on phage. Current Opinion in Biotechnology 6, 73-80.[CrossRef]
    [Google Scholar]
  11. Dore, I., Altschuh, D., Al Moudallal, Z. & Van Regenmortel, M. H. ( 1987; ). Immunochemical studies of tobacco mosaic virus. VII. Use of comparative surface accessibility of residues in antigenically related viruses for delineating epitopes recognized by monoclonal antibodies. Molecular Immunology 24, 1351-1358.[CrossRef]
    [Google Scholar]
  12. Dore, I., Weiss, E., Altschuh, D. & Van Regenmortel, M. H. ( 1988; ). Visualization by electron microscopy of the location of tobacco mosaic virus epitopes reacting with monoclonal antibodies in enzyme immunoassay. Virology 162, 279-289.[CrossRef]
    [Google Scholar]
  13. Fack, F., Hugle-Dorr, B., Song, D., Queitsch, I., Petersen, G. & Bautz, E. K. ( 1997; ). Epitope mapping by phage display: random versus gene-fragment libraries. Journal of Immunology Methods 206, 43-52.[CrossRef]
    [Google Scholar]
  14. Felici, F., Castagnoli, L., Musacchio, A., Jappelli, R. & Cesareni, G. ( 1991; ). Selection of antibody ligands from a large library of oligopeptides expressed on a multivalent exposition vector. Journal of Molecular Biology 222, 301-310.[CrossRef]
    [Google Scholar]
  15. Fischer, R., Schumann, D., Zimmermann, S., Drossard, J., Sack, M. & Schillberg, S. ( 1999; ). Expression and characterization of bispecific single-chain Fv fragments produced in transgenic plants. European Journal of Biochemistry 262, 810-816.[CrossRef]
    [Google Scholar]
  16. Geysen, H. M., Meloen, R. H. & Barteling, S. J. ( 1984; ). Use of peptide synthesis to probe viral antigens for epitopes to a resolution of a single amino acid. Proceedings of the National Academy of Sciences, USA 81, 3998-4002.[CrossRef]
    [Google Scholar]
  17. Gupta, S., Arora, K., Sampath, A., Khurana, S., Singh, S. S., Gupta, A. & Chaudhary, V. K. ( 1999; ). Simplified gene-fragment phage display system for epitope mapping. Biotechniques 27, 328-334.
    [Google Scholar]
  18. Hämmerling, G. J. & Hämmerling, U. ( 1981; ). Production of antibody-producing hybridomas in the rodent system. Research Monographs in Immunology 3, 563-587.
    [Google Scholar]
  19. Jacobsson, K. & Frykberg, L. ( 1995; ). Cloning of ligand-binding domains of bacterial receptors by phage display. Biotechniques 18, 878-885.
    [Google Scholar]
  20. Jacobsson, K. & Frykberg, L. ( 1996; ). Phage display shot-gun cloning of ligand-binding domains of prokaryotic receptors approaches 100% correct clones. Biotechniques 20, 1070-1081.
    [Google Scholar]
  21. Jacobsson, K., Jonsson, H., Lindmark, H., Guss, B., Lindberg, M. & Frykberg, L. ( 1997; ). Shot-gun phage display mapping of two streptococcal cell-surface proteins. Microbiological Research 152, 121-128.[CrossRef]
    [Google Scholar]
  22. Lane, D. P. & Stephen, C. W. ( 1993; ). Epitope mapping using bacteriophage peptide libraries. Current Opinion in Immunology 5, 268-271.[CrossRef]
    [Google Scholar]
  23. Luzzago, A., Felici, F., Tramontano, A., Pessi, A. & Cortese, R. ( 1993; ). Mimicking of discontinuous epitopes by phage-displayed peptides. I. Epitope mapping of human H ferritin using a phage library of constrained peptides. Gene 128, 51-57.[CrossRef]
    [Google Scholar]
  24. Schillberg, S., Zimmermann, S., Voss, A. & Fischer, R. ( 1999; ). Apoplastic and cytosolic expression of full-size antibodies and antibody fragments in Nicotiana tabacum. Transgenic Research 8, 255-263.[CrossRef]
    [Google Scholar]
  25. Scott, J. K. & Smith, G. P. ( 1990; ). Searching for peptide ligands with an epitope library. Science 249, 386-390.[CrossRef]
    [Google Scholar]
  26. Sibille, P. & Strosberg, A. D. ( 1997; ). A FIV epitope defined by a phage peptide library screened with a monoclonal anti-FIV antibody. Immunology Letters 59, 133-137.[CrossRef]
    [Google Scholar]
  27. Smith, G. P. ( 1991; ). Surface presentation of protein epitopes using bacteriophage expression systems. Current Opinion in Biotechnology 2, 668-673.[CrossRef]
    [Google Scholar]
  28. Stephen, C. W., Helminen, P. & Lane, D. P. ( 1995; ). Characterisation of epitopes on human p53 using phage-displayed peptide libraries: insights into antibody–peptide interactions. Journal of Molecular Biology 248, 58-78.[CrossRef]
    [Google Scholar]
  29. Tavladoraki, P., Benvenuto, E., Trinca, S., De Martinis, D., Cattaneo, A. & Galeffi, P. ( 1993; ). Transgenic plants expressing a functional single-chain Fv antibody are specifically protected from virus attack. Nature 366, 469-472.[CrossRef]
    [Google Scholar]
  30. Valadon, P. & Scharff, M. D. ( 1996; ). Enhancement of ELISAs for screening peptides in epitope phage display libraries. Journal of Immunology Methods 197, 171-179.[CrossRef]
    [Google Scholar]
  31. Van Regenmortel, M. H. V. ( 1981; ). Tobamoviruses. In Handbook of Plant Virus Infections and Comparative Diagnosis , pp. 541-564. Edited by E. Kurstak. Amsterdam:Elsevier/North Holland.
  32. Van Regenmortel, M. H. ( 1999; ). The antigenicity of tobacco mosaic virus. Philosophical Transactions of the Royal Society of London Series B 354, 559-568.[CrossRef]
    [Google Scholar]
  33. Verch, T., Yusibov, V. & Koprowski, H. ( 1998; ). Expression and assembly of a full-length monoclonal antibody in plants using a plant virus vector. Journal of Immunology Methods 220, 69-75.[CrossRef]
    [Google Scholar]
  34. Vieira, J. & Messing, J. ( 1987; ). Production of single-stranded plasmid DNA. Methods in Enzymology 153, 3-11.
    [Google Scholar]
  35. Westerwoudt, R. J. ( 1985; ). Improved fusion methods. IV. Technical aspects. Journal of Immunology Methods 77, 181-196.[CrossRef]
    [Google Scholar]
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