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Journal of Biomolecular Screening
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High-Throughput Screening with HyperCyt® Flow Cytometry to Detect Small Molecule Formylpeptide Receptor Ligands

Susan M. Young

Cytometry and Department of Pathology, Cancer Research and Treatment Center, University of New Mexico, Health Sciences Center

Cristian Bologa

Department of Cell Biology and Physiology, Cancer Research and Treatment Center, University of New Mexico, Health Sciences Center

Eric R. Prossnitz

Office of Biocomputing, University of New Mexico, Health Sciences Center

Tudor I. Oprea

Department of Cell Biology and Physiology, Cancer Research and Treatment Center, University of New Mexico, Health Sciences Center

Larry A. Sklar

Bruce S. Edwards

Cytometry and Department of Pathology, Cancer Research and Treatment Center, University of New Mexico, Health Sciences Center

High-throughput flow cytometry (HTFC), enabled by faster automated sample processing, represents a promising high- content approach for compound library screening. HyperCyt® is a recently developed automated HTFC analysis system by which cell samples are rapidly aspirated from microplate wells and delivered to the flow cytometer. The formylpeptide receptor (FPR) family of G protein–coupled receptors contributes to the localization and activation of tissue-damaging leukocytes at sites of chronic inflammation. Here, the authors describe development and application of an HTFC screening approach to detect potential anti-inflammatory compounds that block ligand binding to FPR. Using a homogeneous no-wash assay, samples were routinely processed at 1.5 s/well (~2500 cells analyzed/sample), allowing a 96-well plate to be processed in less than 2.5 min. Assay sensitivity and accuracy were validated by detection of a previously documented active compound with relatively low FPR affinity (sulfinpyrazone, inhibition constant [Ki]=14 µM) from among a collection of 880 compounds in the Prestwick Chemical Library. The HyperCyt® system was therefore demonstrated to be a robust, sensitive, and highly quantitative method with which to screen lead compound libraries in a 96-well format.

Key Words: drug discovery • flow cytometry • automation • ligand binding assay • fluorescence

Journal of Biomolecular Screening, Vol. 10, No. 4, 374-382 (2005)
DOI: 10.1177/1087057105274532
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