Figure 5.

Characteristics of NLC_v2 with Enhanced RNA Loading Capacity

(A) Particle size as measured by dynamic light scattering. Data are reported as Z-average. (B) Denaturing RNA agarose gel electrophoresis of untreated rvRNA (lane 2) and NLC_v2 rvRNA (lane 4) or RNase-treated rvRNA (lane 3) and NLC_v2 rvRNA (lane 5). Data is representative of three independent trials. (C) Percent of RNA bound to NLC_v1 or NLC_v2 as measured by densitometry analysis following denaturing agarose gel electrophoresis of NLC_v1 or NLC_v2 complexed with increasing concentrations of rvRNA. Data is representative of three independent experiments. (D) SEAP expression in BHK cell supernatant 24 hr after incubation with NLC_v1 or NLC_v2 complexed at various N:P ratios with SEAP rvRNA. Data is represented as mean ± SD of three biological replicates and is representative of results from three independent experiments. NLC_v1 data reproduced from Figure 2E for comparison purposes. (E) Guinea pigs (n = 4/group) were immunized with a single 50-μg dose of unformulated ZIKV rvRNA, 5 or 0.5 μg of NLC_v1-formulated ZIKV rvRNA, or with 50, 5, or 0.5 μg of NLC_v2-formulated ZIKV rvRNA via the IM or ID routes, and serum-neutralizing antibody titers, as measured by PRNT₈₀, were assayed 28 days later. Each data point is reported as well as mean ± SD. Experiment performed once. Log₁₀ transform of data in (E) were analyzed by one-way ANOVA with Tukey’s multiple comparison test (5-μg doses between NLC_v1 and NLC_v2 formulations via IM, *p = 0.05, or ID, *p = 0.04).