| Assay Method Information | |
| | Cytopathic Effect Assay (CPE Assay) |
| Description: | MDCK cells (Madin-Daby canine kidney cells) were seeded in a 384-well plate with 6000 cells per well and cultured at 37° C. under 5% CO2 condition overnight. Next day, cell culture medium was exchanged with fresh medium containing relevant concentrations of test compounds and virus Influenza B virus-B/Lee/40 (the titer was 30 TCID50/well). The highest tested concentration of the compounds were 50 μM and then the compounds were diluted by 3-fold serially for a total of 11 concentrations. A cytotoxicity test group was set under the same experimental condition as described above, except that the cell culture medium of cytotoxicity test group didn't contain influenza virus; while a virus control group without drug and a no virus infecting cell control group without drug were set at the same time. Each group was set in duplicate, and incubated at 37° C. under 5% CO2 condition for 3 days. The cell activity was detected by using Promega CellTiter Cell Viability kits, and the data were used for calculating the antiviral effect and cytotoxicity against virus-infected cell of the compound. Data were analyzed by using GraphPad Prism software, and the CPE inhibition ratio and Cell survival ratio were calculated. EC50 value was obtained according to the curve fitting. |
| Affinity data for this assay | |
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