High throughput screening technologies for direct cyclic AMP measurement.

Authors: Gabriel D, Vernier M, Pfeifer MJ, Dasen B, Tenaillon L, Bouhelal R.
Publisher/Year: Assay Drug Dev Technol. 2003 Apr;1(2):291-303.
Pub Med ID/Journal ID: PMID:15090194


A study comparing five different cAMP detection technologies in terms of sensitivity, robustness, and feasibility for HTS is presented. In this report, the following methods are described: a nonhomogeneous DELFIA, and the homogeneous methods based on time-resolved fluorescence (HTRF), luminescent singlet oxygen channeling or ALPHAScreen, FP, and high-affinity enzyme complementation. DELFIA had the highest sensitivity, whereas ALPHAScreen and HTRF shared several advantages, including high sensitivity, broad dynamic range, and minimal reagent addition steps. For G(s)-coupled antagonist screens, we found HTRF and ALPHAScreen the more sensitive and HTS-compatible techniques.