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. 1984 Jul;81(13):3998–4002. doi: 10.1073/pnas.81.13.3998

Use of peptide synthesis to probe viral antigens for epitopes to a resolution of a single amino acid.

H M Geysen, R H Meloen, S J Barteling
PMCID: PMC345355  PMID: 6204335

Abstract

A procedure is described for rapid concurrent synthesis on solid supports of hundreds of peptides, of sufficient purity to react in an enzyme-linked immunosorbent assay. Interaction of synthesized peptides with antibodies is then easily detected without removing them from the support. In this manner an immunogenic epitope of the immunologically important coat protein of foot-and-mouth disease virus (type O1) is located with a resolution of seven amino acids, corresponding to amino acids 146-152 of that protein. Then, a complete replacement set of peptides in which all 20 amino acids were substituted in turn at every position within the epitope was synthesized, and the particular amino acids conferring specificity for the reaction with antibody were determined. It was found that the leucine residues at positions 148 and 151 were essential for reaction with antisera raised against intact virus. A lesser contribution was derived from the glutamine and alanine residues at positions 149 and 152, respectively. Aside from the practical significance for locating and examining epitopes at high resolution, these findings may lead to better understanding of the basis of antigen-antibody interaction and antibody specificity.

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Selected References

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

  1. Atassi M. Z. Antigenic structure of myoglobin: the complete immunochemical anatomy of a protein and conclusions relating to antigenic structures of proteins. Immunochemistry. 1975 May;12(5):423–438. doi: 10.1016/0019-2791(75)90010-5. [DOI] [PubMed] [Google Scholar]
  2. Atassi M. Z. Precise determination of the entire antigenic structure of lysozyme: molecular features of protein antigenic structures and potential of "surface-simulation" synthesis--a powerful new concept for protein binding sites. Immunochemistry. 1978 Dec;15(12):909–936. doi: 10.1016/0161-5890(78)90126-8. [DOI] [PubMed] [Google Scholar]
  3. Barteling S. J., Wagenaar F., Gielkens A. L. The positively charged structural virus protein (VP1) of foot-and-mouth disease virus (type O1) contains a highly basic part which may be involved in early virus-cell interaction. J Gen Virol. 1982 Oct;62(Pt 2):357–361. doi: 10.1099/0022-1317-62-2-357. [DOI] [PubMed] [Google Scholar]
  4. Benjamini E., Michaeli D., Young J. D. Antigenic determinants of proteins of defined sequences. Curr Top Microbiol Immunol. 1972;58:85–134. doi: 10.1007/978-3-642-65357-5_3. [DOI] [PubMed] [Google Scholar]
  5. Bittle J. L., Houghten R. A., Alexander H., Shinnick T. M., Sutcliffe J. G., Lerner R. A., Rowlands D. J., Brown F. Protection against foot-and-mouth disease by immunization with a chemically synthesized peptide predicted from the viral nucleotide sequence. Nature. 1982 Jul 1;298(5869):30–33. doi: 10.1038/298030a0. [DOI] [PubMed] [Google Scholar]
  6. Dreesman G. R., Sanchez Y., Ionescu-Matiu I., Sparrow J. T., Six H. R., Peterson D. L., Hollinger F. B., Melnick J. L. Antibody to hepatitis B surface antigen after a single inoculation of uncoupled synthetic HBsAg peptides. Nature. 1982 Jan 14;295(5845):158–160. doi: 10.1038/295158a0. [DOI] [PubMed] [Google Scholar]
  7. Green N., Alexander H., Olson A., Alexander S., Shinnick T. M., Sutcliffe J. G., Lerner R. A. Immunogenic structure of the influenza virus hemagglutinin. Cell. 1982 Mar;28(3):477–487. doi: 10.1016/0092-8674(82)90202-1. [DOI] [PubMed] [Google Scholar]
  8. Hurrell J. G., Smith J. A., Leach S. J. The detection of five antigenically reactive regions in the soybean leghemoglobin a molecule. Immunochemistry. 1978 May;15(5):297–302. doi: 10.1016/0161-5890(78)90089-5. [DOI] [PubMed] [Google Scholar]
  9. Kazim A. L., Atassi M. Z. A novel and comprehensive synthetic approach for the elucidation of protein antigenic structures. Determination of the full antigenic profile of the alpha-chain of human haemoglobin. Biochem J. 1980 Oct 1;191(1):261–264. doi: 10.1042/bj1910261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kleid D. G., Yansura D., Small B., Dowbenko D., Moore D. M., Grubman M. J., McKercher P. D., Morgan D. O., Robertson B. H., Bachrach H. L. Cloned viral protein vaccine for foot-and-mouth disease: responses in cattle and swine. Science. 1981 Dec 4;214(4525):1125–1129. doi: 10.1126/science.6272395. [DOI] [PubMed] [Google Scholar]
  11. Kurz C., Forss S., Küpper H., Strohmaier K., Schaller H. Nucleotide sequence and corresponding amino acid sequence of the gene for the major antigen of foot and mouth disease virus. Nucleic Acids Res. 1981 Apr 24;9(8):1919–1931. doi: 10.1093/nar/9.8.1919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lerner R. A., Green N., Alexander H., Liu F. T., Sutcliffe J. G., Shinnick T. M. Chemically synthesized peptides predicted from the nucleotide sequence of the hepatitis B virus genome elicit antibodies reactive with the native envelope protein of Dane particles. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3403–3407. doi: 10.1073/pnas.78.6.3403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Meloen R. H., Briaire J. A study of the cross-reacting antigens on the intact foot-and-mouth disease virus and its 12S Subunits with antisera against the structural proteins. J Gen Virol. 1980 Nov;51(Pt 1):107–116. doi: 10.1099/0022-1317-51-1-107. [DOI] [PubMed] [Google Scholar]
  14. Neurath A. R., Kent S. B., Strick N. Specificity of antibodies elicited by a synthetic peptide having a sequence in common with a fragment of a virus protein, the hepatitis B surface antigen. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7871–7875. doi: 10.1073/pnas.79.24.7871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pfaff E., Mussgay M., Böhm H. O., Schulz G. E., Schaller H. Antibodies against a preselected peptide recognize and neutralize foot and mouth disease virus. EMBO J. 1982;1(7):869–874. doi: 10.1002/j.1460-2075.1982.tb01262.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Prince A. M., Ikram H., Hopp T. P. Hepatitis B virus vaccine: identification of HBsAg/a and HBsAg/d but not HBsAg/y subtype antigenic determinants on a synthetic immunogenic peptide. Proc Natl Acad Sci U S A. 1982 Jan;79(2):579–582. doi: 10.1073/pnas.79.2.579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rowlands D. J., Clarke B. E., Carroll A. R., Brown F., Nicholson B. H., Bittle J. L., Houghten R. A., Lerner R. A. Chemical basis of antigenic variation in foot-and-mouth disease virus. Nature. 1983 Dec 15;306(5944):694–697. doi: 10.1038/306694a0. [DOI] [PubMed] [Google Scholar]
  18. Smith J. A., Hurrell J. G., Leach S. J. A novel method for delineating antigenic determinants: peptide synthesis and radioimmunoassay using the same solid support. Immunochemistry. 1977 Aug;14(8):565–568. doi: 10.1016/0019-2791(77)90150-1. [DOI] [PubMed] [Google Scholar]

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