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P2040
A Novel Incubation Procedure to Reduce Edge-Effect in High-Throughput Cell-Based Assays
Presenter Yan Zhai, Eli Lilly and Company, USA
Additional Authors: Karen L. Huss, Karen L. Cox, Weiming Li, Kenneth B. Rank, Bruce A. Wiltermood, Yi Zeng, Susan Jaken, Robert M. Campbell
Cell-based assays are well established as powerful and widely used tools for drug discovery. However because these assays utilize living cells, they can be extremely sensitive to variations in extrinsic chemical and physical parameters. “Edge-effect,” defined as a systematic within-plate column and/or row based bias, is a well established issue encountered in cell-based microplate assay development. The common approach of dealing with edge-effect – simply not using the peripheral wells of the plate – only avoids the problem. To better understand and minimize variations in cell-based assays used in high-throughput screening, we evaluated several parameters and assessed their contribution to well-to-well variation in our 384-well cell-based Luciferase reporter assay. We found that simply placing the cell plates (without lids) inside Nunc 245mm Square BioAssay dishes helped to normalize well-to-well fluctuations in temperature and air-flow within the tissue culture incubator, which resulted in not only reduced well-to-well variation but also a total elimination of assay edge-effect. By implementing this simple and inexpensive approach, we were able to successfully develop and validate a 384-well cell-based reporter assay, and quickly and efficiently screen a 100K compound library.