James J. Devlin

High capacity screening of pooled compounds: Identification of the active compound without re‐assay of pool members

A matrix‐based compound pooling and deconvolution method has been developed that significantly increased the efficiency of a high‐capacity screening program. This method is based on screening pools of 10 compounds. The matrix used to assemble the pools resulted in each compound being assayed twice—each time with a completely different set of pooled compounds. The active compound in an active pool was accurately predicted by determining which compound was present in an active pool both times that a pool containing that compound was tested. This has eliminated the need to re‐assay each individual member of active pools. This approach has been tested with a set of 6,680 compounds in three different assays: inhibition of Factor Xa, inhibition of gastrin‐releasing peptide receptor binding, and inhibition of nitric oxide synthase (isoform II). Thus significant time is saved not only by pooling the compounds for the initial assay but also by avoiding the need to re‐assay all compounds in active pools. Moreover, no false negatives occurred in these assays—an important consideration from a drug discovery perspective.