Fig. 5: CH235.12 encounter state design improves the selection of target mutations.
From: Engineering immunogens that select for specific mutations in HIV broadly neutralizing antibodies
A Aligned structures of the CH505.M5.G458Y trimer bound to either early intermediate CH235.I60 Fab (dark blue gp120, purple VH, dark gray VL) or mature bnAb CH235.12 Fab (dark red gp120, green VH, light gray VL) highlighting the position of loop D residue K279. B (left) Aligned gp120 cores unliganded (light blue; PDB ID 3TGT) and bound (light green; PDB ID 5F96) to CH235.12 (VH orange, VL light gray). (right) View of loop D in the unliganded and CH235.12 bound state showing displacement of residue L277. C Kinetics (left, ka, and middle, kd) and affinity (right) for the CH505.M5.G458Y CE3 design (N386R + V5 disulfide+N197D) with either K279 or N279 (log scaling used for visual clarity). D Mutations frequencies for human heavy (left) and light (right) chain genes in CH235 UCA knock-in mice. Significance was calculated using a two-tailed, non-parametric Mann-Whitney test and is marked by an asterisk (*p < 0.05; Heavy Chain p = 0.056, Light Chain p > 0.999, N = 4 mice per group). E CH235 UCA knock-in mouse serum neutralization titers for CH505 isolate-derived pseudo-viruses. Significance was calculated using a two-tailed, non-parametric Mann-Whitney test and is marked by an asterisk (*p < 0.05; CH505TF N279K p = 0.016, CH505TF N279K + G458Y p = 0.008, N = 4 mice per group). F (left) Representative structure of encounter State 3 highlighting the position of the design mutations (red spheres) and the encounter state contacts (pink spheres) in the CH235.12 HCDR2 proximal sheet. (right) mutation frequencies for key I60 mutations from immunized CH235 UCA knock-in mice. Significance was calculated using a two-tailed, non-parametric Mann-Whitney test and is marked by an asterisk (*p < 0.05; N = 4 mice per group). Source data are provided as a Source Data file.
