Fig. 5
Discovery of early neutralizing intermediates with CDRH3-stabilizing mutations. a CDRH3 alignment for ADS-4 from the Arg97 + disulfide sublineage and GDS-(1–4) from the Gly97 + disulfide sublineage showing the critical CDRH3-stabilizing mutations. Percent nucleotide mutation for the VH gene is shown. b Neutralization capacity for ADS-4 and GDS-(1–4). IC50 values (µg/mL) are shown and indicate a trend in increasing potency and breadth for the Gly97 + disfulide sublineage members. Strain names in bold are autologous and the superinfecting strain (SU) and the week 34 longitudinal time point strains are labeled. Neutralization curves for GDS-1 and GDS-2 antibodies showing weak neutralization of early appearing autologous strains from week 34 are shown. c Binding kinetics of week 34 autologous strain CAP256.3.11.80 to the synthesized antibodies, the UCA and VRC26.25. d Timeline of all tested antibodies from the VRC26 lineage shown for the two major bifurcating branches. All newly identified intermediates for the early stages of the lower major bifurcating branch are in bold. Boxed sequences contain the Cys100a, Cys100q and Arg97 CDRH3 mutations. Only the lower major bifurcating branch leads to the most potent and broad antibodies with stabilizing mutations. Note that VRC26.24, while isolated at week 193, is actually phylogenetically placed on the upper branch by exon analysis and lacks the three stabilizing mutations26
