Fig. 5
From: A human endothelial cell-based recycling assay for screening of FcRn targeted molecules
HERA screening of WT and Fc-engineered hIgG1 variants. a Uptake of WT and Fc-engineered hIgG1 variants at pH 7.4 when 400 nM of each variant was added to the cells followed by 4 h incubation, washing and lysis of the cells. b Recycling of the Fc-engineered hIgG1 variants at pH 7.4 when 400 nM of each variant was added to the cells and incubated for 4 h followed by extensive washing and additional 4 h incubation before sample collection. c The same procedure as in b followed by lysis of the cells. The amounts of hIgG variants in all samples were quantified by ELISA and obtained data are shown as mean ± s.d. of four independent experiments performed in triplicates. ns > 0.05, ****p < 0.0001, by one-way ANOVA (Tukey’s multiple comparison test). Relative d uptake, e recycling and f residual amount calculated from data (a–c), g uptake, h recycling and i residual amounts when WT and the Fc-engineered hIgG1 variants were initially incubated at pH 6.0. The amounts of hIgG variants in all samples were quantified by ELISA, and obtained data are shown as mean ± s.d. of two independent experiments performed in triplicates. ns > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001, by one-way ANOVA (Tukey’s multiple comparison test)
