a C57BL/6 mice were vaccinated two times, with 3 weeks between vaccinations, as in Figure 4a. Dose level of VLP or nano-VLP (nVLP) was 5 µg, based on GP content, and dose level of the adjuvant poly-ICLC was 10 µg. Time to death in days, expressed as percentage survival in each group. All animals except those in the adjuvant-only poly-ICLC control group survived. b Anti-glycoprotein titers of mice from A inoculated with adjuvant alone, or adjuvant plus sucrose-gradient control VLP, frozen nVLP, lyophilized nVLP, and lyophilized nVLP heated to 75°C for 1 h. c Time to death in days, expressed as percentage survival in each group of mice vaccinated with 5 or 20 µg nVLP doses, without adjuvant. The experiment compares results for nVLP stored frozen, lyophilized nVLP, lyophilized and heated nVLP, and a control of sucrose-gradient purified VLP. p-values determined using Fisher’s exact test to compare survival of each treatment group vs. saline. d Anti-glycoprotein titers of mice from c. P values determined using one-tailed Student’s t-test where *p < 0.05, **p < 0.005, ***p < 0.0005. “Lyoph” indicates lyophilized material.