Generation of chimeric hamster polyomavirus VP1 virus-like particles harboring three tumor-associated antigens

We have examined hamster polyomavirus VP1 virus-like particles (HaPyV VP1 VLPs) as potential carriers of tumor-associated antigens (TAAs). Three different HLA-*A–restricted human TAA epitopes: TRP – tyrosinase-related protein-2 epitope (amino acid sequence FVWLHYYSV), MAGE – the MAGE A family protein epitope (LVHPLLLKY) and HTERT – human telomerase reverse transcriptase epitope (ILAKFLHWT), were inserted into VP1 protein into positions #1 (81–88aa), #3 (244–246aa) and #4 (289-294aa) separately and into position #4 as one fused insert produced from all three TAAs. The constructed chimeric proteins were expressed in yeast Saccharomyces cerevisiae and after purification were capable to form VLPs. HaPyV-VP1 appeared as a suitable carrier of multiple TAAs. The insertion of separate hydrophobic peptides into different sites of VP1 VLPs has appeared to be a particularly promising approach, because insertion of a long fused hydrophobic peptide unfavorably influenced the VPL assembly and yield. Abbreviations: HaPyV – hamster polyomavirus, VP1 – virus protein 1, VLPs – virus-like particles, TAA – tumor-associated antigen, CTL – cytotoxic T lymphocyte, TRP – tyrosinase-related protein-2 epitope, MAGE – the MAGE A family protein epitope, HTERT – human telomerase reverse transcriptase epitope