Fig. 3.

Purification, Epitope Binding, and C1q Binding of ZE3 RIC. (A) Following protein G affinity purification of ZE3 C-RICs and N-RICs, samples of the C-RIC and N-RIC elutions were analyzed by SDS-PAGE gel stained with Coomassie (left two panels) and by a western blot (rightmost panel) probed with anti-human IgG + HRP. Abbreviations: R, reducing and boiled conditions, and NR, non-reducing and non-boiled conditions. (B) Various concentrations of purified 6D8 antibody (HL), tagless N-terminally fused ZE3 (ZE3-HL), or tagless N-terminally fused Zika E ectodomain (ZE-HL) were added to ELISA plates coated with 10 μg/ml carrier protein with 6D8 epitope fusion. The bound constructs were detected with goat anti-human kappa HRP-labeled antibody. The OD450 values were plotted on the y-axis while the concentration of the probing antibody was plotted on the x-axis. (C) C1q binding of purified C-RIC, N-RIC, and 6D8 was measured by ELISA. Mean OD450 values from 10-fold serial dilutions starting at 10 μg/ml are shown.