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Journal of Biomolecular Screening
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Correction for Interference by Test Samples in High-Throughput Assays

Adam B. Shapiro

AstraZeneca R&D Boston, Waltham, Massachusetts, adam.shapiro{at}astrazeneca.com

Grant K. Walkup

AstraZeneca R&D Boston, Waltham, Massachusetts

Thomas A. Keating

AstraZeneca R&D Boston, Waltham, Massachusetts

In high-throughput biochemical assays performed in multiwell plates, the effect of test samples on the activity of the biochemical system is usually measured by optical means such as absorbance, fluorescence, luminescence, or scintillation counting. The test sample often causes detection interference when it remains in the well during the measurement. Interference may be due to light absorption, fluorescence quenching, sample fluorescence, chemical interaction of the sample with a detection reagent, or depression of the meniscus. A simple method is described that corrects for such interference well by well. The interference is measured in a separate artifact assay plate. An appropriate arithmetic correction is then applied to the measurement in the corresponding well of the activity assay plate. The correction procedure can be used for single-point screening or potency measurements on serial dilutions of test samples.

Key Words: high-throughput screening • interference • correction • fluorescence resonance energy transfer • scintillation proximity assay

This version was published on September 1, 2009

Journal of Biomolecular Screening, Vol. 14, No. 8, 1008-1016 (2009)
DOI: 10.1177/1087057109341768
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