Measuring Intracellular Calcium Fluxes in High Throughput Mode

Title: Measuring Intracellular Calcium Fluxes in High Throughput Mode

Volume: 6 Issue: 4

Author(s): Chris Chambers, Fiona Smith, Christine Williams, Sandra Marcos, Zhen Han Liu, Paul Hayter, Giuseppe Ciaramella, Wilma Keighley, Phil Gribbon and Andreas Sewing

Affiliation:

Keywords: intracellular calcium fluxes, protein coupled receptors, chimeric, fluorometric imaging plate reader

Abstract: The measurement of intracellular calcium fluxes in real time is widely applied within the pharmaceutical industry to measure the activation of G-protein coupled receptors (GPCRs), either for pharmacological characterisation or to screen for new surrogate ligands. Initially restricted to Gq coupled GPCRs, the introduction of promiscuous and chimeric G-proteins has further widened the application of these assays. The development of new calcium sensitive dyes and assays has provided sensitive, homogeneous assays which can be readily applied to high throughput screening (HTS). In this paper we describe the full automation of this assay type using a fluorometric imaging plate reader (FLIPR™) integrated into a Beckman / Sagian system to establish a simple robotic system that is well suited for the current medium throughput screening in this area of lead discovery. Using a recently completed HTS we discuss important determinants for FLIPR™ based screening, highlight some limitations of the current approach, and look at the requirements for future automated systems capable of keeping up with expanding compound files.

Cite this article as:

Chambers Chris, Smith Fiona, Williams Christine, Marcos Sandra, Liu Han Zhen, Hayter Paul, Ciaramella Giuseppe, Keighley Wilma, Gribbon Phil and Sewing Andreas, Measuring Intracellular Calcium Fluxes in High Throughput Mode, Combinatorial Chemistry & High Throughput Screening 2003; 6 (4) . https://dx.doi.org/10.2174/138620703106298446