High throughput screening technologies for direct cyclic AMP measurement.

Gabriel D, Vernier M, Pfeifer MJ, Dasen B, Tenaillon L, Bouhelal R


Novartis Pharma AG, Basel, Switzerland

Assay Drug Dev Technol. 2003;1(2):291-303.

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 Gs-coupled antagonist screens, we found HTRF® and ALPHAScreen® the more sensitive and HTS-compatible techniques.

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