Orientation of Antigen Display on Self-Assembling Protein Nanoparticles Influences Immunogenicity

Schneider, Cosette G.; Taylor, Justin A.; Sibilo, Michael Q.; Miura, Kazutoyo; Mallory, Katherine L.; Mann, Christopher; Karch, Christopher; Beck, Zoltan; Matyas, Gary R.; Long, Carole A.; Bergmann-Leitner, Elke; Burkhard, Peter; Angov, Evelina

Self-assembling protein nanoparticles (SAPN) serve as a repetitive antigen delivery platform with high-density epitope display; however, antigen characteristics such as size and epitope presentation can influence the immunogenicity of the assembled particle and are aspects to consider for a rationally designed effective vaccine. Here, we characterize the folding and immunogenicity of heterogeneous antigen display by integrating (a) dual-stage antigen SAPN presenting the P. falciparum (Pf) merozoite surface protein 1 subunit, PfMSP1(19), and Pf cell-traversal protein for ookinetes and sporozoites, PfCelTOS, in addition to (b) a homogenous antigen SAPN displaying two copies of PfCelTOS. Mice and rabbits were utilized to evaluate antigen-specific humoral and cellular induction as well as functional antibodies via growth inhibition of the blood-stage parasite. We demonstrate that antigen orientation and folding influence the elicited immune response, and when appropriately designed, SAPN can serve as an adaptable platform for an effective multi-antigen display.