Synthetic peptide epitope‐based polymers: controlling size and determining the efficiency of epitope incorporation

Abstract

The assembly of synthetic peptide-based vaccines that incorporate multiple epitopes is a major goal of vaccine development, because such vaccines will potentially allow the immunization of outbred populations against a number of different pathogens. We have shown that free radical-induced polymerization of individual peptide epitopes results in the incorporation of multiple copies of the same or different epitopes into high molecular weight immunogens (O'Brien-Simpson, N.M., Ede, N.J., Brown, L.E., Swan, J. & Jackson, D.C. (1997) Polymerization of unprotected synthetic peptides: a view toward synthetic peptide vaccines. J. Am. Chem. Soc.119, 1183-1188; Jackson, D.C., O'Brien-Simpson, N., Ede, N.J. & Brown, L.E. (1997) Free radical induced polymerization of synthetic peptides into polymeric immunogens. Vaccine 15, 1697-1705). The ability to control the size of these polymers, to determine the physical and chemical properties of the backbone material and also to know the extent to which individual peptide epitopes are incorporated are important manufacturing considerations and form the subject of this study. We show here that the polymerization process is highly efficient with at least 70% of peptides incorporated into the resulting polymer, that acrylamide and acryloylated amino acids can be used as comonomers with peptide epitopes in the polymerization reaction and that the choice of the comonomer can influence the properties of the resulting polymer. We also show that the size of chain growth polymers is restricted in the presence of chain transfer agents, that the resulting polymer size can be predicted and that there is little or no difference in the immunogenicity of polymers that range in apparent molecular size between 18 kDa and 335 kDa. The successful polymerization of peptide epitopes with an acryloyl-amino acid creates the potential for introducing different physical and chemical properties into artificial protein immunogens.

Effects on Rotavirus Cell Binding and Infection of Monomeric and Polymeric Peptides Containing α2β1 and αxβ2 Integrin Ligand Sequences

“…Peptides were assembled manually with Fmoc chemistry throughout as described previously (44). In order to enable polymerization of individual peptides, an acryloyl group was attached to individual peptides; derivitization of peptides with the acryloyl (CH 2 ϭ CHCO-) group with acryloyl chloride is described elsewhere (42).…”

“…Polymerization was allowed to proceed for 18 h at room temperature. High-molecular-weight material resulting from the polymerization reaction was separated from low-molecular-weight reactants with a column (1.6 cm by 60 cm) of Sephadex G-25 (fractionation range, 1,000 to 5,000 Da) installed in a high-pressure liquid chromatography system as previously described (44). Separations were performed at a flow rate of 4 ml per min with 50 mM ammonium hydrogen carbonate, pH 8.1, as the eluant.…”

A self‐adjuvanting multiepitope immunogen that induces a broadly cross‐reactive antibody to hepatitis C virus†

“…23 Although neutralizing antibody responses are not associated with viral clearance in acute hepatitis C infection, they do have a role in controlling viral replication in patients with chronic hepatitis C. 24 We adopted a strategy incorporating a combinatorial epitope library into a single immunogen that uses novel chemistry to develop vaccine candidates with broad coverage against HCV. [25][26][27][28][29] In this study, we report the design, assembly and immunological properties of selfadjuvanting multiepitope immunogens that produce broad cross-reactive antibodies capable of binding to HCV E1/E2 proteins expressed at the surface of transfected cells that are also capable of capturing different HCV quasispecies circulating in the serum of infected carriers. This approach provides a strategy for the development of a preventative vaccine against HCV.…”

Gonococcal transferrin binding protein chimeras induce bactericidal and growth inhibitory antibodies in mice

“…The NB domain has a predicted molecular mass of ~44 kDa, while the L2 domain is ~9 kDa. The small size of L2 could account for its relatively poor immunogenicity [48]. Second, full-length gonococcal TbpA is not as immunogenic as TbpB [19].…”