Extended Data Fig. 5: Characterization of antibody responses in macaques.

a, Binding of serum from macaques primed with RC1-4fill VLPs. ELISAs of the serum from eight macaques primed with RC1-4fill VLPs and PGT121 to RC1 (black) and RC1-glycanKO (grey) are shown. b, Binding of serum from macaques primed with RC1-4fill VLPs. ELISA of the serum from eight macaques primed with RC1-4fill VLP and one naive macaque to RC1 (black) and the sequentially less modified Env proteins 11MUTB (grey) and 10MUT (white). The human bNAbs PGT121 and 3BNC60 were used as controls at 5 μg ml⁻¹, and the serum was evaluated at a 1:100 dilution and seven additional threefold serial dilutions. c, The affinities (K_D) for RC1 of different macaque antibodies isolated after a prime with VLP-RC1-4fill and the corresponding inferred germline-reverted antibodies as determined by bio-layer interferometry (OCTET). d, Binding of an anti-idiotypic antibody that recognizes the inferred germline of PGT121/10-1074 to monoclonal antibodies isolated from macaques primed with VLP-RC1-4fill. The inferred germline (iGL) of PGT121/10-1074, two chimeric antibodies comprising the mutated (MT) heavy chain (HC) and inferred germline light chain (LC) of PGT121 (PGT121 HCMT-LC iGL) or the inferred germline HC and the mutated LC of PGT121 (PGT121HC iGL-LCMT) and different inferred germline bNAbs were used as controls. a, b, d, Results are shown as the area under the ELISA curve (AUC). e, Comparison of binding mode between the vaccine-elicited antibodies (Ab275_MUR, Ab874_NHP and Ab897_NHP) and the V3-glycan patch bNAbs 10-1074, PGT128 and PGT135. RC1 trimer is shown in grey from above and all Fabs are modelled onto the same trimer. For clarity, only one Fab per trimer is shown. f, Interactions between Ab897_NHP conserved light chain motifs and RC1 gp120. Lime, DNS motif in CDRL3; red, gp120 GDIR; pink, NIG motif in CDRL1; teal, gp120 V1 loop. Each AUC value corresponds to one ELISA curve.