Figure 4
From: Engineering of anti-human interleukin-4 receptor alpha antibodies with potent antagonistic activity
Epitope mapping of anti-IL-4Rα Abs by alanine scanning mutagenesis. (a) Competitive ELISA showing the percentage of bound IL-4Rα (10 and 50 nM) to plate-coated hIL-4-mFc in the presence of the indicated Abs (20, 100, and 500 nM) compared to that without the Ab competitor. Data are represented as mean ± SD (n = 3). Statistical analyses were performed using a two-way ANOVA followed by the Newman-Keuls post-test. *P < 0.05, **P < 0.01, ***P < 0.001; ns, not significant versus dupilumab analogue. (b) Overall structure of the human IL-4Rα:IL-4 complex (PDB: 1IAR). Magnified section shows the residues of IL-4Rα putatively involved in IL-4 binding. (c) The percent relative binding of the indicated hIL-4-mFc (5 nM) and anti-IL-4Rα Abs (2.5 nM of 4 R34 and 100 pM of 4 R34.1, 4 R34.1.19 and dupilumab analogue) to IL-4Rα alanine mutants compared to that of wild-type IL-4Rα. Data are represented as mean ± SD (n = 3). Statistical analyses were performed using a one-way ANOVA followed by the Newman-Keuls post-test. *P < 0.05, **P < 0.01, ***P < 0.001 versus binding to wild-type IL-4Rα.
