Figure 2.

5D5 paratope glycosylation mediates high-affinity binding. (A) Interactions formed by mAb 5D5 H.Asn98-linked GlcNAc moiety and surrounding CDR residues with PfCSP. H-bonds are shown as black dashed lines, and salt bridges are shown as blue dashed lines. (B–F) Binding kinetics of twofold dilutions of 293F (C), 293S (GnT I^−/−; D), Endo H (E), and 5D5Δg (F) Fab glycoform variants to full-length PfCSP. (B) Mean on- and off-rates (k_on and k_off, respectively) of the 5D5 Fab glycoform variants binding to full-length PfCSP are plotted on the left and right y axes, respectively, for three independent experimental replicates. Mean k_on rates are shown as red circles, and mean k_off rates as blue triangles. Error bars represent one SD from the mean. (C–F) Representative sensorgrams are shown in black and 1:1 model best fits in red. Mean K_D values are as listed. K_D values and k_on and k_off rates were determined by FortéBio’s Data Analysis software 9.0. Standard error values are reported as the SD. Data are representative of three independent measurements. Corresponding glycan structures are shown using symbols adhering to the Symbol Nomenclature for Glycans (Varki et al., 2015).