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11/6/606    most recent
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This version was published on September 1, 2006
Journal of Biomolecular Screening, Vol. 11, No. 6, 606-616 (2006)
DOI: 10.1177/1087057106288183

Assay Concordance between SPA and TR-FRET in High-Throughput Screening

Oliver Von Ahsen

Schering AG, Enabling Technologies, Department of Assay Development & High Throughput Screening, Berlin, Germany

Anne Schmidt

Schering AG, Enabling Technologies, Department of Assay Development & High Throughput Screening, Berlin, Germany

Monika Klotz

Schering AG, Enabling Technologies, Department of Assay Development & High Throughput Screening, Berlin, Germany

Karsten Parczyk

Schering AG, Enabling Technologies, Department of Assay Development & High Throughput Screening, Berlin, Germany

High-throughput screening (HTS) of large chemical libraries has become the main source of new lead compounds for drug development. Several specialized detection technologies have been developed to facilitate the cost- and time-efficient screening of millions of compounds. However, concerns have been raised, claiming that different HTS technologies may produce different hits, thus limiting trust in the reliability of HTS data. This study was aimed to investigate the reliability of the authors most frequently used assay techniques: scintillation proximity assay (SPA) and homogeneous time-resolved fluorescence resonance energy transfer (TR-FRET). To investigate the data concordance between these 2 detection technologies, the authors screened a large subset of the Schering compound library consisting of 300,000 compounds for inhibitors of a nonreceptor tyrosine kinase. They chose to set up this study in realistic HTS scale to ensure statistical significance of the results. The findings clearly demonstrate that the choice of detection technology has no significant impact on hit finding, provided that assays are biochemically equivalent. Data concordance is up to 90%. The little differences in hit findings are caused by threshold setting but not by systematic differences between the technologies. The most significant difference between the compared techniques is that in the SPA format, more false-positive primary hits were obtained.

Key Words: assay techniques • tyrosine kinase • high-throughput screening • TR-FRET • SPA • assay concordance


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