FIG 6 .

Analysis of the antibody response to the N332 supersite in mouse antisera. (A) Schematic view of the immunization regimen to test two N332 nanoparticles in BALB/c mice (W, week). (B) ELISA binding of mouse antisera from the 1KIG_L_ES-2-FR group to 3 pairs of monomeric and particulate N332 scaffolds (M, treated mouse; N, naïve mouse). (C) ELISA binding of mouse antisera from the 1KIG_L_ES-2-FR group to ferritin, a mutated 1GUT_A_ES antigen—1GUT_A_ES_Mut3—with three glycan knockout mutations (N295A, N301A, and N332A), and the native-like gp140.664.R1 trimer (46). (D) Schematic view of the immunization regimen to test the gp140.664.R1 trimer and a trimer-prime/epitope-boost strategy in BALB/c mice. (E) ELISA binding of mouse antisera to the gp140.664.R1 trimer, a pair of monomeric and particulate N332 scaffolds, a V1V2 nanoparticle, and a gp120 nanoparticle (48) for 4 subjects from the trimer-alone group. (F) ELISA binding of mouse antisera to the same set of antigens as described for panel E for 4 subjects from the heterologous prime/boost group. The immunogens and the regimen details are labeled on the diagram for panels A and D. A naïve mouse sample was included in the ELISA as a control, as shown in panels B and E. The EC₅₀ values are labeled for all ELISA plots in panels B, C, E, and F, except for instances in which the highest OD₄₅₀ value was below 0.1 or in the cases of ambiguous data fitting.