Figure 2.
Design and biochemical characterization of 3nv.2 SOSIP. (A) Design of 426c-based triple immunogen to present CD4bs (blue), V3 (green), and V2 (purple) epitopes. (B) Schematics of single (IGT2), double (IGT2–RC1), and triple (3nv.2) immunogen constructs used in this study. (C and D) 3nv.2 and 3nv.1 triple immunogen characterization by (C) SEC and (D) SDS-PAGE. (E) Top: 2D class averages demonstrating 3nv.2 is predominantly trimeric. Bottom: 6.6-Å single-particle 3nv.2 cryo-EM density map. (F) Schematic for the generation of SOSIP–mi3 nanoparticles using the SpyCatcher–SpyTag system. (G) Characterization of purified SOSIP–mi3 nanoparticles by SDS-PAGE. R, reduced; NR, non-reduced; SC, SpyCatcher. (H) Negative-stain EM of SOSIP–mi3 nanoparticles. Scale bar = 100 nm.

Design and biochemical characterization of 3nv.2 SOSIP. (A) Design of 426c-based triple immunogen to present CD4bs (blue), V3 (green), and V2 (purple) epitopes. (B) Schematics of single (IGT2), double (IGT2–RC1), and triple (3nv.2) immunogen constructs used in this study. (C and D) 3nv.2 and 3nv.1 triple immunogen characterization by (C) SEC and (D) SDS-PAGE. (E) Top: 2D class averages demonstrating 3nv.2 is predominantly trimeric. Bottom: 6.6-Å single-particle 3nv.2 cryo-EM density map. (F) Schematic for the generation of SOSIP–mi3 nanoparticles using the SpyCatcher–SpyTag system. (G) Characterization of purified SOSIP–mi3 nanoparticles by SDS-PAGE. R, reduced; NR, non-reduced; SC, SpyCatcher. (H) Negative-stain EM of SOSIP–mi3 nanoparticles. Scale bar = 100 nm.

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