High resolution cryo-EM structures of HIV SOSIP Env with antibodies. (A) Naturally-elicited antibodies in complex with HIV SOSIP Env spike: From left to right (first row), CD4 binding site Ab VRC01 (EMD-40281), glycan cluster Ab 2G12 (EMD-20224), V1/V2-apex directed Ab PGT145 (EMD-29288). From left to right (second row), V3-glycan targeted Ab DH270.6 (EMD-20818), gp120-gp41 interface Ab 8ANC195 (EMD-0485), fusion peptide-targeting Ab VRC34 (EMD-28617), membrane proximal external region targeted Ab 4E10 (EMD-25045). (B) Vaccine-elicited antibodies in complex with HIV SOSIP Env spike: From left to right (first row), V3-glycan mouse Ab MU89 (EMD-27706), CD4 binding site macaque Ab DH1285 (EMD-41823), FP-targeting human Ab 2C06 (EMD-29725), MPER targeted human Ab DH1317.4 (EMD-44246). (C) Cryo-EM based polyclonal epitope mapping (cryo-EMPEM): From left to right (first row), polyclonal antibodies were selected from rhesus macaques Rh.32034 (Grp1): pAbC-1(C3/V5 targeted, EMD-23223) and pAbC-2(N241/N289 glycan hole targeted, EMD-23224); From left to right (second row) Rh.33104 (Grp1): pAbC-1(N241/N289 glycan hole targeted, EMD-23227) and pAbC-2(N241/N289 glycan hole targeted, EMD-23228); From left to right (third row) Rh.33311 (Grp2): pAbC-1(FP targeted, EMD-23236), pAbC-2(N611-glycan targeted, EMD-23237), and pAbC-5(C3/V5 targeted, EMD-23240). Antibody heavy chain in medium purple and light chain in pink, Env gp120 in dark gray and gp41 in light steel blue throughout Figure 3. Each panel includes a schematic representation (created with BioRender.com).