| Assay Method Information | |
| | Coronavirus Antiviral Assay |
| Description: | The human β-coronavirus strain OC43 was purchased from ATCC (Manassas, Va.; item numbers VR-1558 and VR-740, respectively). 24 hours prior to dosing, HeLa human cervix epithelial cells (ATCC, CCL-2) or MRC-5 human lung fibroblast (ATCC, CCL-171) were plated in 96 well plates at a density of 1.5×105/ml in DMEM medium supplemented with 10% fetal bovine serum, 1% HEPES buffer, 1% Penicillin/Streptomycin and 1% non-essential amino acids (all Mediatech, Manassas, Va.). At the day of infection, serially diluted compounds were added to cells and incubated for 4 hours. After the end of the 4 hour pre-incubation period, cells were infected with either coronavirus strain OC43 or 229E. The virus inoculum was selected to cause 80-90% cytopathic effect. Infected cells were incubated for five days at 37° C., 5% CO2. To develop the assay, 100 μl media was replaced with 100 μl CellTiter-Glo reagent (Promega, Madison, Wis.), and incubated for 10 min at room temperature. Luminescence was measured on a Victor X3 multi-label plate reader. Potential compound cytotoxicity was determined using uninfected parallel cultures. |
| Affinity data for this assay | |
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