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A Novel High-Throughput Screening Assay for Sickle Cell Disease Drug Discovery

Department of Physiology and Biophysics, Keck School of Medicine, University of Southern California, Los Angeles, Childrens Hospital, Los Angeles, California

John S. Cambridge

Department of Physiology and Biophysics, Keck School of Medicine, University of Southern California, Los Angeles

Cage S. Johnson

Department of Medicine, Keck School of Medicine, University of Southern California, Los Angeles

Herbert J. Meiselman

Department of Physiology and Biophysics, Keck School of Medicine, University of Southern California, Los Angeles

Timothy C. Fisher

Department of Physiology and Biophysics, Keck School of Medicine, University of Southern California, Los Angeles

Tamas Alexy

Department of Physiology and Biophysics, Keck School of Medicine, University of Southern California, Los Angeles, alexy{at}usc.edu

Although the pathophysiology and molecular basis of sickle cell disease (SCD) were described more than half a century ago, an effective and safe therapy is not yet available. This may be explained by the lack of a suitable high-throughput technique that allows rapid screening of thousands of compounds for their antisickling effect. The authors have thus developed a novel high-throughput screening (HTS) assay based on detecting the ability of red blood cells (RBC) to traverse a column of tightly packed Sephacryl chromatography beads. When deoxygenated, sickle RBC are rigid and remain on the top of the column. However, when deoxygenated and treated with an effective antisickling agent, erythrocytes move through the Sephacryl media and produce a red dot on the bottom of the assay tubes. This approach has been adapted to wells in a 384-well microplate. Results can be obtained by optical scanning: The size of the red dot is proportional to the antisickling effect of the test molecule. The new assay is simple, inexpensive, reproducible, requires no special reagents, and should be readily adaptable to robotic HTS systems. It has the potential to identify novel drug candidates, allowing the development of new therapeutic options for individuals affected with SCD. (Journal of Biomolecular Screening. 2009:330-336)

Key Words: sickle cell disease • high-throughput screening • cell-based assay • assay development • drug discovery

Journal of Biomolecular Screening, Vol. 14, No. 4, 330-336 (2009)
DOI: 10.1177/1087057109333975


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