Testing antivirals in the cell-based ELISA: factors that influence the potency of drugs

Authors

• RA Gross
• E Postnikova
• Y Cong
• LE Dewald
• BJ Hart
• J Dyall
• H Zhou
• N Deiuliis
• J Michelotti
• A Honko
• RS Bennett
• MR Holbrook
• GG Olinger
• LE Hensley
• PB Jahrling

Abstract

From 2013-2016, West Africa experienced the largest Ebola virus (EBOV) outbreak in history. During this time, there was an urgent need to quickly screen existing drugs for anti-EBOV activity in order to potentially repurpose them for immediate, off label, compassionate use. Various high-throughput cell-based assays have been developed for use in anti-EBOV screens; however, no study or report have shown how varying conditions within screening assays can influence the apparent potency of a drug. This study demonstrates the effects of different parameters (cell types, virus input-multiplicity of infection, assay infection endpoint, drug exposure time, and fetal bovine serum (FBS) concentration) on drug potency in a cell-based EBOV-ELISA assay using an antibody against viral protein 40 (VP40). Drug potency was evaluated by a drugs half maximal effective concentration (EC50). Signal-to-noise was evaluated when determining which parameters give the optimal conditions for improving drug potency. These results demonstrate that different assay conditions will affect the EC50 values obtained for the same drug. Of the parameters tested, it was determined, cell type, assay endpoint, and virus input were the most significant. These results further demonstrate a need for consistent standard operating procedures while performing cell-based assays to generate reproducible and reliable data on the antiviral activity of drugs.

Scientific Focus Area: Virology

This page was last updated on Friday, March 26, 2021