Figure 6. Asn276 glycosylation frequency modulated VRC01_GL-class IgGs recognition of the 426c core.

(A–D) BLI binding data and associated K_D values of 426c core constructs, expressed in HEK293 GnTI^-/- cells, with two immobilized VRC01_GL-class IgGs. VRC01_GL IgG binding was assessed against the 426c core (A) and the 426c S278T core (B). 12A21_GL binding to the 426c core (C) and 426c S278T core (D) were also tested. (E–H) BLI binding data and corresponding K_D values of 426c core constructs, expressed using HEK293 GnTI^-/- cells and treated with EndoH, with VRC01_GL-class IgGs. VRC01_GL IgG binding was assessed against the EndoH-treated 426c core (E) and the 426c S278T core (F). 12A21_GL interactions with the EndoH-treated 426c core (G) and 426c S278T core (H) were also tested. The concentrations of 426c core injected and the color key are indicated on each panel. Fit curves are colored as black dotted lines. The vertical dotted lines indicate the transition between association and dissociation phases.

Figure 6—figure supplement 1. VRC01_GL binding affinity was not significantly affected by the identity of the residue 278 in the absence of a glycan at position Asn276.

(A) BLI kinetics parameters determined from panels (B–G) and Figure 6. (B–D) BLI binding data and determined equilibrium dissociation constants for VRC01_GL IgG binding to 426c S278A core (A), 426c S278V core (B), and 426c S278R core. (E–G) BLI binding data and determined equilibrium dissociation constants of 12A21_GL IgG binding to 426c S278A core (A), 426c S278V core (B), and 426c S278R core. The concentrations of 426c core injected are indicated on each panel. Fit curves are colored black. The vertical dashed lines indicate the transition between association and dissociation phases.