FIG 7.
Compound KD for ZIKV RdRp determined by initial fluorescence analysis. The protein was applied at a final concentration of 100 nM. A twofold dilution series (16 concentration points) starting from 500 μM was prepared for each compound. The competition between Cpd15 and Cpd17 was performed in a similar way with a fixed concentration of 100 μM Cpd17 with a twofold dilution series of Cpd15 starting from 500 μM. Samples were filled into hydrophilic capillaries (NanoTemper Technologies) and incubated shortly before measurement at 37°C using a Monolith NT.LabelFree instrument (NanoTemper Technologies). The curves were generated using MO.Affinity software by NanoTemper Technologies. The value of the initial fluorescence was plotted against the ligand concentration to derive the dose-response curve. The KD values were obtained with the initial fluorescence mode, measuring the fluorescence intensity variation upon compound binding. The inset table shows the data processing statistics exported from MO.Affinity software.