Assay Method Information

Assay Name:  plaque reduction assay
Description:  Confluent monolayers of Vero E6 ACE2 cells in 12-well plates were washed once with DMEM and infected with approximately 50 plaque forming units (PFUs) of SARS-CoV-2 in each well. The plates were incubated on a rocker at 37 C for 45 min for virus adsorption. The virus inoculum was removed and replaced by overlay medium (DMEM containing 1% low-melting point agarose without serum) containing 3-fold serial dilutions of the test compounds and placed in a 37 C CO2 incubator until plaques could be visualized under light. The cells were then fixed with a 4% formaldehyde solution for at least 30 min, and the overlaid agarose was removed and stained with a 0.2% (w/v) crystal violet solution. The plaques were counted by visual examination, and the required concentration to reduce the 50% plaque number (EC50) was calculated relative to the control without test compounds.
Affinity data for this assay
 

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