Fig. 3: Characterization and immunogenicity of an E2E1-NP based on a genotype 3 strain.
a SEC profiles from a Superose 6 column of AMS3a E2E1-I53-50A and AMS3a E2E1-NPs. b NS-EM images and 2D class averages of AMS3a E2E1-NPs. The white scale bar represents 200 nm. c Rabbits were immunized at weeks 0 and 4 with E2, E2E1-I53-50A, and E2E1-NP, all based on the AMS3a strain. Antibody responses were measured at week 6. d Binding titers to E2E1-foldon (AMS3a) and e HVR1 peptide (AMS3a). f Competition ELISA of immunized rabbit sera. Residual binding of mAbs was measured after incubating rabbit sera with AMS3a-derived E1E2 from cell lysates. Each dot represents the value of three independent experiments measured in duplo or triplo. Bars indicate the median values. g Neutralization titers against the sequence-matched AMS3a HCVpp. h Global geometric mean titers (GGMT) were calculated based on the neutralization ID50 titers against fifteen non-matched HCVpp strains. Each dot represents a single rabbit serum. Neutralization ID50 titers of the individual viruses are depicted in Table S1. i Breadth of the neutralizing response defined as the number of HCVpp strains neutralized with an ID50 titer above 40. j Correlations between breadth of the serum response (i) and residual binding measured in competition ELISA (f). Spearman r and p-value (two-tailed) are indicated. Horizontal lines in (d), (e), (g)–(i) indicate the medians values. Significant differences between groups in (d)–(i) were determined using Kruskal–Wallis test followed by Dunn’s post-test, n = 6 rabbit sera per group. P-values for significant differences are indicated on top of the graphs. Source data are provided as a Source data file.
