Wang Yuren, Jones Philip
Neuroscience Discovery Research, Wyeth Research, Princeton, NJ, USA.
Methods Mol Biol. 2009;572:247-59. doi: 10.1007/978-1-60761-244-5_16.
Scintillation proximity assay (SPA) is a homogenous and versatile technology for the simple and sensitive detection of the interaction of protein targets with their ligands. Herein, we described a SPA assay developed to identify compounds that bind to human fatty acid amide hydrolase (FAAH). This SPA assay utilizes the specific binding of [(3)H]-R(+)-methanandamide ((3)H-MAEA), a competitive nonhydrolyzed FAAH inhibitor, to FAAH expressing microsomes and evaluates its displacement by FAAH inhibitors. In contrast to the classical SPA radioligand binding assay which detects bound ligand, in our assay the released radiolabel is detected through its interaction with the SPA beads. This novel SPA assay has been validated and demonstrated to be simple, sensitive, and amenable to high-throughput screening.
闪烁邻近分析（SPA）是一种用于简单且灵敏地检测蛋白质靶点与其配体相互作用的均相通用技术。在此，我们描述了一种开发用于鉴定与人脂肪酸酰胺水解酶（FAAH）结合的化合物的SPA分析方法。该SPA分析利用[(3)H]-R(+)-甲磺酰胺（(3)H-MAEA）（一种竞争性非水解FAAH抑制剂）与表达FAAH的微粒体的特异性结合，并评估FAAH抑制剂对其的置换作用。与检测结合配体的经典SPA放射性配体结合分析不同，在我们的分析中，通过释放的放射性标记与SPA微球的相互作用来检测它。这种新型SPA分析方法已经得到验证，并证明其简单、灵敏且适用于高通量筛选。
