Correction for interference by test samples in high-throughput assays.
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.
Shapiro
Adam B
AB
AstraZeneca R&D Boston, Waltham, Massachusetts 02451, USA. adam.shapiro@astrazeneca.com
Walkup
Grant K
GK
Keating
Thomas A
TA
D000042:Absorption / Q000502:physiology; D000465:Algorithms; D016477:Artifacts; D002138:Calibration; D003627:Data Interpretation, Statistical; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods / Q000592:standards; D057166:High-Throughput Screening Assays / Q000379:methods* / Q000592:standards*; D020128:Inhibitory Concentration 50; D008027:Light; D008163:Luminescent Measurements / Q000379:methods / Q000592:standards; D015588:Observer Variation; D012107:Research Design / Q000592:standards; D012588:Scintillation Counting / Q000379:methods / Q000592:standards; D013048:Specimen Handling / Q000379:methods / Q000592:standards