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).
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Walter, G. (1986) Production and use of antibodies against synthetic peptides. J. Immunol. Methods 88, 149–161.
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.
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.
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.
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.
Dryberg, T. and Oldstone, M. B. A. (1986) Peptides as antigens. J. Exp. Med. 164, 1344–1349.
Ponsati, B., Giraldt, E., and Andreu, D. (1989) A synthetic strategy for simultaneous purification-congujation of antigenic peptides. Anal. Biochem. 181, 389–395.
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.
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.
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.
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.
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.
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.
Reichlin, M. (1980) Use of glutaraldehyde as a coupling agent for proteins and peptides. Methods Enzymol. 70, 159–165.
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.
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.
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© 1996 Humana Press Inc., Totowa, NJ
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Carter, J.M. (1996). Conjugation of Peptides to Carrier Proteins via Glutaraldehyde. In: Walker, J.M. (eds) The Protein Protocols Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1007/978-1-60327-259-9_117
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DOI: https://doi.org/10.1007/978-1-60327-259-9_117
Publisher Name: Humana Press
Print ISBN: 978-0-89603-338-2
Online ISBN: 978-1-60327-259-9
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