View All Posters

P2007
Improvements in Data Quality & Assay Performance Using Aequorin for Calcium Based Screening.
Presenter Jane Miller, GlaxoSmithKline, United Kingdom
Additional Authors: Stephen Brough
In pursuit of faster, more accurate and more precise pharmacological screening for reduced cost, we have carried out an extensive analysis into the methods we use for characterising G-protein coupled receptor (GPCR) coupled increases in intracellular calcium. In the last decade we have relied almost exclusively on fluorescent dye-based technologies to measure calcium mobilisation. The aequorin technology was developed in the 1990s as a calcium sensitive bioluminescent protein for use in GPCR assays, but the advent of new luminescence detectors and screening platforms has renewed interest in using this system in the high-throughput screening environment. We describe here the relative benefits observed from implementing an aequorin- based detection method for screening the activity of GPCRs in both 384 and 1536 well format in comparison to traditional fluorescent dye-based technologies. Details of benefits observed for both structure activity relationship (SAR) and single shot high-throughput screens (HTS) are included in this analysis. Improvements in data quality, fidelity and reproducibility; simplification in assay protocols and processes; cost comparisons for both HTS and SAR screening campaigns as well as intangible benefits from switching between formats are shown here. In conclusion, we demonstrate significant improvements in data quality, which has lead to a reduction in assay failure. This, in combination with the savings seen in both costs and time, along with the scope for further development as the technology becomes embedded, has enabled us to establish aequorin-based calcium mobilisation assays as the standard in our laboratory