ameon: a comparative study of fluorescence lifetime and ADP-Glo assays for protein kinase screening

Drug discovery requires reliable and consistent compound profiling processes to ensure robustness during the lead identification phase. Significant improvements in assay reagents (FLEXYTE, Almac) and introduction of an advanced HTS-compatible microplate reader (ameon, TTP Labtech) position fluorescence lifetime at the forefront of cost-effective screening technologies. Utilisation of the FLEX17 long lifetime fluorophore enables discrimination from interfering
compounds present in compound libraries. In turn, the ameon reader applies a proprietary data recording technology to
deliver precise lifetime measurement at high throughput for easy integration into screening workflows.

Conventional assays based on luminescent and fluorescent readouts have issues with quenching, auto-fluorescence or
light sensitivity of reagents which in turn cause false positives and negatives. In this study, the FLEXYTE lifetime and ADPGlo Kinase (Promega) assays were compared as primary screening technologies by addressing a validation set of compounds (6,334 in total, AstraZeneca) for a serine/threonine protein kinase target. Hits identified from
primary screening (10 M) were confirmed by concentrationresponse (CR). All primary hits (including those not confirmed by ADP-Glo) were subsequently assessed in an orthogonal mass spectrometry kinase assay (LCMS).

Download: lifetime_vs_ADP_Glo_kinase_screeningAZ_TTPLabtech_poster_SLAS2016.pdf