Gabriel Daniela, Vernier Magali, Pfeifer Martin J, Dasen Boris, Tenaillon Laurent, Bouhelal Rochdi
Novartis Pharma AG, Lead Discovery Center, Basel, Switzerland.
Assay Drug Dev Technol. 2003 Apr;1(2):291-303. doi: 10.1089/15406580360545107.
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.
本文介绍了一项针对五种不同的cAMP检测技术在灵敏度、稳健性及用于高通量筛选（HTS）的可行性方面的比较研究。在本报告中，描述了以下方法：非均相解离增强镧系元素荧光免疫分析（DELFIA），以及基于时间分辨荧光（HTRF）、发光单线态氧通道或AlphaScreen、荧光偏振（FP）和高亲和力酶互补的均相方法。DELFIA具有最高的灵敏度，而AlphaScreen和HTRF具有若干共同优点，包括高灵敏度、宽动态范围和最少的试剂添加步骤。对于G(s)偶联拮抗剂筛选，我们发现HTRF和AlphaScreen是更灵敏且与高通量筛选兼容的技术。
