Evaluation of dynamic mass redistribution technology for pharmacological studies of recombinant and endogenously expressed g protein-coupled receptors

Assay Drug Dev Technol. 2008 Feb;6(1):83-94. doi: 10.1089/adt.2007.126.

Abstract

The Epic cell assay technology (Corning Inc., Corning, NY) uses a resonant waveguide grating optical biosensor to measure cellular response to ligands manifested through dynamic mass redistribution (DMR) of cellular contents. The DMR measurement is a noninvasive, label-free assay that can be used to assess the pharmacological properties of compounds. In this study, a panel of 12 compounds was evaluated against two G protein-coupled receptor (GPCR) targets in recombinant expressed cell lines using the Corning Epic system in 384-well microplates. The evaluation was performed in a double-blinded fashion such that the identity and properties of both the GPCR targets and compounds were unknown to the researchers at the time of the study. Analysis of the DMR response from cell stimulation was used to identify compounds that functioned as agonists or antagonists and to evaluate the associated efficacy and potency. DMR results were shown to have good agreement with data obtained from cyclic AMP and calcium flux assays for compounds evaluated. A further analysis was performed and successfully identified the signaling pathways that the two GPCRs activated. In addition, the DMR measurement was able to detect responses from an endogenous receptor in these cells. The Epic DMR technology provides a generic platform amenable to pharmacological evaluation of cellular responses to GPCR activation in a label-free live cell assay format.

MeSH terms

  • Animals
  • Biosensing Techniques
  • CHO Cells
  • Calcium / metabolism
  • Cricetinae
  • Cricetulus
  • Cyclic AMP / metabolism
  • Drug Evaluation, Preclinical / methods*
  • Drug Evaluation, Preclinical / statistics & numerical data*
  • Kinetics
  • Models, Statistical
  • Receptors, G-Protein-Coupled / biosynthesis
  • Receptors, G-Protein-Coupled / drug effects*
  • Receptors, G-Protein-Coupled / genetics
  • Recombinant Proteins
  • Signal Transduction / drug effects

Substances

  • Receptors, G-Protein-Coupled
  • Recombinant Proteins
  • Cyclic AMP
  • Calcium