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Conjugation of Peptides to Carrier Proteins via Glutaraldehyde

  • J. Mark Carter
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

There are three common purposes for conjugation of peptides. The most common is induction of humoral immunity (1). This is the production of antibodies capable of binding to the peptide immunogen. The antibodies are elaborated by plasma cells, which are terminally differentiated B-lymphocytes. However, in order for immunity to be successfully induced in a secondary anamnestic response, the immunogen must also react with T-lymphocytes. Many peptides contain B-cell epitopes, but not T-cell epitopes. (Such molecules are called haptens.) Coupling these molecules to a large carrier protein containing T-cell epitopes allows the induction of a B-cell response to the entire immunogen, including the peptide. New synthetic peptides thus offer promise as vaccines (2).

Keywords

Carrier Protein Lipid Emulsion Keyhole Limpet Hemocyanin Peptide Molecule Capture Antigen 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Walter, G. (1986) Production and use of antibodies against synthetic peptides. J. Immunol. Methods 88, 149–161.PubMedCrossRefGoogle Scholar
  2. 2.
    Patarroyo, M. E., Amador, R., Clavijo, P., Moreno, A., Guzman, F., Romero, P., Tascon, R., Franco, A., Murillo, L. A., Ponton, G., and Trujillo, G. (1988) A synthetic vaccine protects humans against challenge with asexual blood stages of Plasmodium falciparum malaria. Nature 332, 158–161.PubMedCrossRefGoogle Scholar
  3. 3.
    Rubenstein, K. E., Schneider, R. S., and Ullman, E. F. (1971) Homogenous enzyme immunoassay, a new immunochemical technique. Biophys. Biochem. Res. Commun. 47, 846–851.CrossRefGoogle Scholar
  4. 4.
    VanRegenmortel, M. H. V., Briand, J. P., Muller, S., and Plaue, S. (1988) Laboratory Techniques in Biochemistry and Molecular Biology, vol. 19 (Burdon, R. H. and Van Knippenberg, P. H., eds.), Elsevier, Amsterdam.Google Scholar
  5. 5.
    Lerner, R. A., Green N., Alexander, H., Liu, F.-T., Sutcliffe, J. G., and Shinnick, T. M. (1981) 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. USA 78, 3403–3407.PubMedCrossRefGoogle Scholar
  6. 6.
    Dryberg, T. and Oldstone, M. B. A. (1986) Peptides as antigens. J. Exp. Med. 164, 1344–1349.CrossRefGoogle Scholar
  7. 7.
    Ponsati, B., Giraldt, E., and Andreu, D. (1989) A synthetic strategy for simultaneous purification-congujation of antigenic peptides. Anal. Biochem. 181, 389–395.PubMedCrossRefGoogle Scholar
  8. 8.
    Satterthwait, A. C., Arrhenius, T., Hagopian, R. A., Zavala, F., Nussenzweig, V., and Lerner, R. A. (1988) Conformational restriction of peptidyl immunogens with covalent replacements for the hydrogen bond. Vaccine 6, 99–103.PubMedCrossRefGoogle Scholar
  9. 9.
    Alving, C. R., Richards, R. L., Moss, J., Alving, L. I., Clements, J. D., Shiba, T., Kotani, S., Wirtz, R. A., and Hockmeyer, W. T. (1986) Effectiveness of liposomes as potential carriers of vaccines: applications to cholera toxin and human malaria sporozoite antigen. Vaccine 4, 166–172.PubMedCrossRefGoogle Scholar
  10. 10.
    Schaaper, W. M. M., Lankohof, H., Pujik, W. C., and Meleon, R. H. (1989) Manipulation of antipeptide immune response by varying the coupling of the peptide with the carrier protein. Mol. Immunol. 26, 81–85.PubMedCrossRefGoogle Scholar
  11. 11.
    Tsao, J., Lin, X., Lackland, H., Tous, G., Wu, Y., and Stein, S. (1991) Internally standardized amino acid analysis for determining peptide/carrier protein coupling ratio. Anal. Biochem. 197, 137–142.PubMedCrossRefGoogle Scholar
  12. 12.
    Kolodny, N. and Robey, F. A. (1990) Conjugation of synthetic peptides to proteins: quantitation from S-carboxymethylcysteine released upon acid hydrolysis. Anal. Biochem. 187, 136–140.PubMedCrossRefGoogle Scholar
  13. 13.
    Peeters, J. M., Hazendonk, T. G., Beuvery, E. C., and Tesser, G. I. (1989) Comparison of four bifunctional reagents for coupling peptides to proteins and the effect of the three moieties on the immunogenicity of the conjugates. J. Immunol. Methods 120, 133–143.PubMedCrossRefGoogle Scholar
  14. 14.
    Reichlin, M. (1980) Use of glutaraldehyde as a coupling agent for proteins and peptides. Methods Enzymol. 70, 159–165.PubMedCrossRefGoogle Scholar
  15. 15.
    Kirkeby, S., Jakobsen, P., and Moe, D. (1987) Glutaraldehyde—“pure and impure.” A spectroscopic investigation of two commercial glutaraldehyde solutions and their reaction products with amino acids. Anal. Lett. 20(2), 303–315.Google Scholar
  16. 16.
    Baron, M. H. and Baltimore, D. (1982) Antibodies against the chemically synthesized genome-Linked protein of poliovirus react with native virus-specific proteins. Cell 28, 395–404.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1996

Authors and Affiliations

  • J. Mark Carter
    • 1
  1. 1.CytogenPrinceton

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