Abstract
Bioconjugates play an important role in several fields of biomolecular and biomedicinal sciences. Protein/polypeptide-based conjugates with covalently attached epitope peptides are considered as potential synthetic vaccine candidates and/or target antigens in affinity-based bioassays. This chapter describes the synthesis of two- and three-component bioconjugates using water-soluble branched chain polymericpolypeptides with multiple amino and/or carboxyl groups as macromolecular partners and oligopeptides as epitopes with small molecular mass. The synthetic procedures outline three major strategies for the incorporation of multiple copies of uniformly oriented peptide epitopes. In the first example, chloroacetylated polypeptide is conjugated with SH-peptide to form a thioether linkage. Second, two independent oligopeptides are introduced into a macromolecule by amide and disulfide bonds, respectively. In the third example, a new procedure is reported for the formation of disulfide bridges by the use of Npys-modified polypeptide and SH-peptide.
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References
Hudecz, F. (2001) Manipulation of epitope function by modification of peptide structure: a minireview. Biologicals 29, 197–207.
Zeng, W., Ghosh, S., Macris, M., Pagnon, J., and Jackson, D. C. (2001) Assembly of synthetic peptide vaccines by chemoselective ligation of epitopes: influence of different chemical linkages and epitope orientations on biological activity. Vaccine 19, 3843–3852.
Mezö, G., Mezö, I., Pimm, et al. (1996) Synthesis, conformation, biodistribution and hormon related in vitro antitumor effect of GnRH antagonist branched poly peptide conjugate. Bioconjugate Chemistry 7, 642–650.
Hudecz, F., Kóczán, G., and Reményi, J. (2003) Peptide or protein based delivery and targeting, in Molecular Pathomechanisms and New Trends in Drug Research (Keri, G. and Toth, I., eds.), Taylor and Francis Group, London, pp. 553–578.
Hudecz, F., Reményi, J., Szabó, R., et al. (2003) Drug targeting by macromol ecules without recognition unit? J. Mol. Recognition 16, 288–298.
Hudecz, F. and Szekerke, M. (1980) Investigation of drug-protein interactions and the drug-carrier concept by the use of branched polypeptides as model sys tems. Synthesis and characterization of the model peptides. Coll. Czech. Chem. Commun. 45, 933–940.
Mezö, G., Kajtár, J., Nagy, L, Szekerke, M., and Hudecz, F. (1997) Carrier design: Synthesis and conformational studies of poly[L-lysine] based branched polypep tides with hydroxyl groups. Biopolymers 42, 719–730.
Hudecz, F., Pimm, M. V., Rajnavölgyi, É., et al. (1999) Carrier design: New generation of polycationic branched polypeptides containing OH groups with prolonged blood survival and diminished in vitro cytotoxicity. Bioconjugate Chemistry 10, 781–790.
Hudecz, F. and Price, M. R. (1992) Monoclonal antibody binding to peptide epi topes conjugated to synthetic branched polypeptide carriers. Influence of the car rier upon antibody recognition. J. Immunol. Methods 147, 201–210.
Wilkinson, K. A., Vordermeier, M. H., Wilkinson, R., Iványi, J., and Hudecz, F. (1998) Synthesis and in vitro T cell immunogenicity of conjugates with dual speci ficities: attachment of epitope peptides of 16 kDa and 38 kDa proteins from M. tuberculosis to branched polypeptide. Bioconjugate Chemistry 9, 539–547.
Vordermeier, H. M., Harris, D. P., Roman, E., Lathigra, R., Moreno, C., and Ivanyi, J. (1991) Identification of T-cell stimulatory peptides from the 38 kDa protein of M. tuberculosis. J. Immunol. 147, 1023–1029.
Friscia, G., Vordermeier, H. M., Pasvol, G., Harris, D. P., Moreno, C., and Iványi, J. (1995) Human T cell responses to peptide epitopes of the 16-kD antigen in tuberculosis. Clin. Exp. Immunol. 102, 53–57.
Wilkinson, K. A., Hudecz, F., Vordermeier, H. M., Iványi, J., and Wilkinson, R. J. (1999) Enhancement of the T cell response to a mycobacterial peptide by conju gation to synthetic branched polypeptide. Eur. J. Immunol. 29, 2788–2796.
Hilbert, Á., Hudecz, F., Mez, G., et al. (1994) The influence of branched polypeptide carriers on the immunogenicity of predicted epitopes of HS V-1 glycoprotein D. Scand. J. Immunol. 40, 609–617.
Mezö, G., Dalmadi, B., Mucsi, I., Bösze, S., Rajnavölgyi, É., and Hudecz, F. (2002) Peptide based vaccine design: Synthesis and immunological characterisa tion of branched polypeptide conjugates comprising the 276-284 immunodom inant epitope of HSV-1 glycoprotein D. J. Peptide Science 8, 107–117.
Hudecz, F., Nagy, I. B., Kóczán, G., Alsina, M. A., and Reig, F. (2001) Carrier design: influence of charge on interaction of branched polymeric polypeptides with phospholipid model membranes, in Biomedical Polymers and Polymer Therapeutics (Chiellini, E., Sunamoto, J., Migliaresi, C., Ottenbrite, R. M., and Cohn, D. eds.), Kluwer Academic/Plenum Publishers, New York, pp. 103–120.
IUPAC-IUB Commission on Biochemical Nomenclature. (1972) Biochem. J. 127, 753–756.
IUPAC-IUB Commission on Biochemical Nomenclature. (1984) Eur. J. Biochem. 138, 9–37.
Hudecz, F., Kovács, P., Kutassi-Kovács, S., and Kajtár, J. (1984) GPC, CD and sedimentation analysis of poly-Lys and branched chain poly-Lys-poly-DL-Ala polypeptides. Colloid Polym. Sci. 262, 208–212.
Mezö, G., de Oliveira, E., Krikorian, D., et al. (2003) Synthesis and comparision of antibody recognition of conjugates containing herpes simplex virus type 1 glycoprotein D epitope VII. Bioconjugate Chemistry 14, 1260–1269.
Van der Ploeg, J. R., Drijfhout, J. W., Feijlbrief, M., Bloemhoff, W., Welling, G. W., and Welling-Wester, S. (1989) Immunological properties of multiple repeats of a linear epitope of herpes simplex virus type 1 glycoprotein D. J. Immunol. Methods 124,211–217.
Carfi, A., Willis, S. H., Whitbeck, J. C., et al. (2001) Herpes simplex virus glyco protein D bound to the human receptor HveA. Molecular Cell 8, 169–179.
Carlsson, J., Drevin, H., and Axen, R. (1978) Protein thiolation and reversible protein-protein conjugation. N-Succinimidyl 3-(2-pyridyldithio) propionate, a new heterobifunctional reagent. Biochem. J. 173, 723–728.
Matsueda, R. and Walter, R. (1980) 3-nitro-2-piridinesulfenyl (Npys) group. Int. J. Peptide Protein Res. 16, 392–401.
Mezñ, G., Mihala, N., Andreu, D., and Hudecz, F. (2000) Conjugation of epitope peptides to branched chain polypeptides via Cys(Npys). Bioconjugate Chemistry 11, 484–491.
Hudecz, F., Hilbert, Á., Mezñ, G., et al. (1993) Epitope mapping of 273-284 region of HSV glycoprotein D by synthetic branched polypeptide carrier conjugates. Peptide Res. 6, 263–271.
Stuchbury, T., Shipton, M., Norris, R., et al. (1975) A reporter group delivery system with both absolute and selevtive specificity for thiol groups and an improved fluorescent probe containing the 7-nitrobenzo-2-oxa-1,3-diazole moiety. Biochem. J. 151, 417–432.
Kaiser, E., Colescott, R. L., Bossinger, C. D., and Cook, P. I. (1970) Color test for detection of free terminal amino groups in the solid-phase synthesis of peptides. Anal. Biochem. 34, 595–598.
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© 2005 Humana Press Inc., Totowa, NJ
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Hudecz, F. (2005). 13 Synthesis of Peptide Bioconjugates. In: Howl, J. (eds) Peptide Synthesis and Applications. Methods in Molecular Biology™, vol 298. Humana Press. https://doi.org/10.1385/1-59259-877-3:209
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DOI: https://doi.org/10.1385/1-59259-877-3:209
Publisher Name: Humana Press
Print ISBN: 978-1-58829-317-6
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