Fig. 7
L234F and A327G in the IgG1/4 Fc CH2 domain control cellular responses to Fc hexamers via altering their affinity for specific FcγR. a DoE analysis of FcγRIIIb binding. Binding of Alexafluor-647-labeled DoE Fc hexamers to HEK cells expressing FcγRIIIb was measured by flow cytometry. DoE analysis was performed using the mean binding at 3, 1, and 0.3 μg/ml Fc hexamer. Results are displayed as an effects plot with data representing the difference in arithmetic mean from IgG1 WT. Error bars represent 95% confidence intervals. b Single mutant IgG1 Fc hexamer L234F and A327G binding to neutrophils in whole blood. Data represent the mean ± SD of n = 2 donors. c Single mutant IgG1 Fc hexamer L234F and A327G binding to platelets. Data represent the mean ± SEM of n = 3 donors heterozygous for R/H131 FcγRIIa polymorphism. d, e Surface plasmon resonance analysis of L234F and A327G IgG1 Fc hexamer single mutants. Equilibrium binding response of Fc hexamers binding to d FcγRIIIb and e FcγRIIa
