Abstract
Cell-based beta-lactamase reporter gene assays designed to measure the functional responses of G-protein-coupled receptors (GPCRs) were miniaturized to less than 2 microL total assay volume in a 3456-well microplate. Studies were done to evaluate both receptor agonists and antagonists. The pharmacology of agonists and antagonists for target GPCRs originally developed in a 96-well format was recapitulated in a 3456-well microplate format without compromising data quality or EC(50)/IC(50) precision. These assays were employed in high-throughput screening campaigns, allowing the testing of more than 150,000 compounds in 8 h. The instrumentation used and practical aspects of the assay development are discussed.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Biological Assay / instrumentation*
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Biological Assay / methods*
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Combinatorial Chemistry Techniques
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Cricetinae
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Dose-Response Relationship, Drug
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Drug Evaluation, Preclinical / instrumentation
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Drug Evaluation, Preclinical / methods*
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Genes, Reporter
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Humans
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Inhibitory Concentration 50
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Nanotechnology / methods
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Receptors, G-Protein-Coupled / agonists*
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Receptors, G-Protein-Coupled / antagonists & inhibitors*
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Receptors, Neurotensin / drug effects
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Receptors, Neurotensin / genetics
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Receptors, Oxytocin / agonists
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Receptors, Oxytocin / antagonists & inhibitors
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Receptors, Oxytocin / genetics
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Reproducibility of Results
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Spectrometry, Fluorescence
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beta-Lactamases / genetics
Substances
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Receptors, G-Protein-Coupled
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Receptors, Neurotensin
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Receptors, Oxytocin
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beta-Lactamases