Presentation: P03043

GPCR’s are of significant interest to the pharmaceutical industry. As a consequence there is a clear need to generate robust, 'off the shelf' functional screening strategies. GTPγS assays are often problematic to develop due to the dependence of different receptor/G-protein pairs on unique combinations of assay components. As such there has been great demand for an assay development and optimization strategy for GTPγS assays. To address this issue we have utilized Modde (Umetrics) statistical software, automated assay optimization software (Beckman Coulter) and a Biomek 2000 Laboratory Automation Workstation (Beckman Coulter) to test a large number of different assay conditions, varying the concentrations of GDP, MgCl₂, NaCl, Saponin and DTT. This approach of parallel rather than sequential testing of conditions has consistently developed more robust GTPγS assays. An example of the process and its application will be presented.

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