Miniaturization of whole live cell-based GPCR assays using microdispensing and detection systems

J Biomol Screen. 2004 Apr;9(3):186-95. doi: 10.1177/1087057103260070.

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

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Assay / instrumentation*
  • Biological Assay / methods*
  • Combinatorial Chemistry Techniques
  • Cricetinae
  • Dose-Response Relationship, Drug
  • Drug Evaluation, Preclinical / instrumentation
  • Drug Evaluation, Preclinical / methods*
  • Genes, Reporter
  • Humans
  • Inhibitory Concentration 50
  • Nanotechnology / methods
  • Receptors, G-Protein-Coupled / agonists*
  • Receptors, G-Protein-Coupled / antagonists & inhibitors*
  • Receptors, Neurotensin / drug effects
  • Receptors, Neurotensin / genetics
  • Receptors, Oxytocin / agonists
  • Receptors, Oxytocin / antagonists & inhibitors
  • Receptors, Oxytocin / genetics
  • Reproducibility of Results
  • Spectrometry, Fluorescence
  • beta-Lactamases / genetics

Substances

  • Receptors, G-Protein-Coupled
  • Receptors, Neurotensin
  • Receptors, Oxytocin
  • beta-Lactamases