| Assay Method Information | |
| | HTRF Binding Assay |
| Description: | Asymmetric bis-benzimidazole compounds (BBI) of the invention were assessed in a biochemical homogeneous time resolved fluorescence (HTRF) binding assay adapted from the human STING WT binding assay (“HTRF, A guide to Homogeneous Time Resolved Fluorescence”, (2021) PerkinElmer Cisbio; Mathis, G. Clinical Chemistry, 41(9):1391-1397). Briefly, 6His-tagged STING protein was incubated with terbium cryptate-labeled anti-6His antibody, d2-labeled 2′,3′-cGAMP, and varying concentrations of test articles in a 384-well plate format. Donor and acceptor emission signals were measured for each well by plate reader at 665 and 615 nm, respectively, and the ratio of the signals used to calculate percent inhibition of d2-labeled 2′,3′-cGAMP, cyclic dinucleotide binding. Dose-response curves generated from these data were used to calculate 1C50 values. |
| Affinity data for this assay | |
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