Extended Data Fig. 3: Antibody responses in wild-type mice.

a, ELISA cross-reactivity of serum from RC1-immunized wild-type mice to 11MUTB. Binding of the serum from wild-type mice primed with RC1 to RC1, 11MUTB, 10MUT and BG505 is shown in blue. Binding of the human bNAbs 10-1074 (green) and 3BNC117 (red) was evaluated at 5 μg ml⁻¹ as a control. n = 2. b, FACS plots showing the gating strategy to isolate single RC1⁺RC1-glycanKO⁻ germinal centre B cells (DUMP⁻ (CD4⁻, CD8⁻, F4/80⁻, NK1.1⁻, CD11b⁻, CD11c⁻ and Gr-1⁻) B220⁺CD95⁺GL7⁺RC1-glycanKO⁻RC1⁺) from the spleen and draining lymph nodes of wild-type mice primed with RC1 or RC1-4fill. c, Binding of the mouse antibodies Ab275_MUR and Ab276_MUR to a V3 loop-consensus C peptide (see Methods). Human antibodies 3869 and 3074 were used as positive controls. Antibodies were evaluated at 30 μg ml⁻¹. n = 2. d, Representative sensograms from two independent SPR-binding experiments of Ab275_MUR Fab injected over immobilized RC1 (left) or 11MUTB (right). Experimental binding curves (red) are overlaid with predicted curves (black) derived from a 1:1 binding model. Representative of 3 independent experiments. e, Binding of Ab276_MUR and its inferred germline version (Ab276_MURGL) to RC1 (left) and RC1-glycanKO (right) at 30 μg ml⁻¹. The human monoclonal antibodies PGT121 (green) and 3BNC60 (red) were used as controls at 5 μg ml⁻¹.