3nv.2 binds to iGLs targeting three bNAb epitopes. (A) SPR sensorgrams of SOSIP–mi3 nanoparticles injected over bNAb iGLs at a concentration of ∼0.5 mg/ml. First row: The CD4bs-specific single immunogen IGT2 only binds to CD4bs bNAb precursors (IOMA iGL, left), and not V3 (10-1074/PGT121 iGL, middle) or V2 (PG9 or PG16 iGL, right) bNAb precursors. Second row: Incorporating V3-targeting mutations into IGT2 creates an immunogen that binds CD4bs and V3 bNAb precursors, but not V2 bNAb precursors. Third row: Incorporating V3- and V2-targeting residues into the CD4bs-targeting IGT2 SOSIP creates an immunogen (3nv.2) that binds to the CD4bs (IOMA iGL, left), V3 (10-1074/PGT121 iGL, middle), and V2 (PG9 or PG16 iGL, right) bNAb precursors. Fourth row: The parental 426c Env does not bind to any of the bNAb precursors. (B) 3nv.2 SOSIP injected in a dilution series over bNAb iGLs starting at top concentrations of 10 µM or 5 µM as indicated. 3nv.2 SOSIP binds to multiple CD4bs precursors, including IOMA iGL (left), BG24 iGL (middle), and VRC01 iGL (right). (C) 3nv.2 SOSIP injected in a dilution series over bNAb UCAs and an iGL starting at a top concentration of 10 µM. 3nv.2 SOSIP binds to multiple V3 precursors, including BG18 iGL (left), DH270 UCA (middle), and BF520 UCA (right). Representative sensorgrams are from at least two independent experiments. UCA, unmutated common ancestor; RU, resonance unit.
Figure 3.

3nv.2 binds to iGLs targeting three bNAb epitopes. (A) SPR sensorgrams of SOSIP–mi3 nanoparticles injected over bNAb iGLs at a concentration of ∼0.5 mg/ml. First row: The CD4bs-specific single immunogen IGT2 only binds to CD4bs bNAb precursors (IOMA iGL, left), and not V3 (10-1074/PGT121 iGL, middle) or V2 (PG9 or PG16 iGL, right) bNAb precursors. Second row: Incorporating V3-targeting mutations into IGT2 creates an immunogen that binds CD4bs and V3 bNAb precursors, but not V2 bNAb precursors. Third row: Incorporating V3- and V2-targeting residues into the CD4bs-targeting IGT2 SOSIP creates an immunogen (3nv.2) that binds to the CD4bs (IOMA iGL, left), V3 (10-1074/PGT121 iGL, middle), and V2 (PG9 or PG16 iGL, right) bNAb precursors. Fourth row: The parental 426c Env does not bind to any of the bNAb precursors. (B) 3nv.2 SOSIP injected in a dilution series over bNAb iGLs starting at top concentrations of 10 µM or 5 µM as indicated. 3nv.2 SOSIP binds to multiple CD4bs precursors, including IOMA iGL (left), BG24 iGL (middle), and VRC01 iGL (right). (C) 3nv.2 SOSIP injected in a dilution series over bNAb UCAs and an iGL starting at a top concentration of 10 µM. 3nv.2 SOSIP binds to multiple V3 precursors, including BG18 iGL (left), DH270 UCA (middle), and BF520 UCA (right). Representative sensorgrams are from at least two independent experiments. UCA, unmutated common ancestor; RU, resonance unit.

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