FIGURE 6.

Hemagglutinin diffusion in varied hydrogel formations. (a) Mice were vaccinated subcutaneously with a single administration of HA (5 μg) in different formulations of PNP hydrogels comprising TLR7/8a‐NPs or an alum bolus control. Serum antibodies were collected and analyzed via ELISA at the indicated timepoints. All experimental gel groups performed significantly better than the bolus Alum control (b‐d; N = 5). (b) (i) Total IgG endpoint titers over time following immunization with HA entrapped in various hydrogel formulations comprising TLR7/8a‐NPs (10% valency; 20 μg total TLR7/8a dose) indicating that these slow release gels yield more potent and durable antibody responses than Alum. (ii) AUC calculation of endpoint titers over time indicated that 2:10, 1:10, and 2:5 hydrogel formulations all performed similarly, while the 1:5 hydrogel formulation exhibited lower total potency. (c) (i) Total IgG endpoint titers over time and (ii) AUC of endpoint titer over time following immunization with HA entrapped in gel formulations comprising various TLR7/8a‐NP valency (10% or 50% with a 20 μg total TLR7/8a dose). (d) (i) Total IgG endpoint titers over time and (ii) AUC of endpoint titer over time following immunization with HA entrapped in PNP hydrogels comprising various doses of TLR7/8a‐NP (10% valency; total TLR7/8a doses equal to 50, 20, 10, or 5 μg) demonstrating no significant difference in antibody response with TLR7/8a dose