临近闪烁分析法在高通量筛选中的应用

Scintillation Proximity Assays in High Throughput Screening

起源于放射性免疫分析的临近闪烁分析法(scintillation proximity assay,SPA)是一种均相、灵敏、快速和简便的基于闪烁载体的分析平台。该平台可用于筛选药物靶点的先导化合物和研究其生理过程。由于无需分离,易于固定药物靶点和检测其活性,SPA成为一种重要的高通量筛选方法。由于放射性标记分子和亲和标签分子的多样化和商业化、以及液闪计数器和液相操作等技术的发展,SPA已经广泛用于受体结合、高通量药物筛选、酶分析、放射性免疫分析、蛋白质-蛋白质相互作用和细胞水平分析等方面。本文阐述了SPA原理,讨论了其关键技术(包括闪烁载体、液闪计数器和放射性标记分子),分析了其评价体系;同时简述了SPA分析的发展,并介绍了其在高通量筛选中的应用实例,归纳了存在的问题,给出了未来的发展趋势。目前,基于SPA和荧光分析方法已成为高通量药物筛选的热点研究领域,这些筛选技术的革新必然提升我们对细胞体系生物学的全面理解和促进先导化合物筛选过程的显著进步。

Originated from a radioimmunoassay method, scintillation proximity assay （SPA） is a homogenous, sensitive, fast and simple scintillant carrier-based platform. This platform can be used to screen lead compounds for drug targets and investigate their biological processes. SPA becomes an important method in high throughput screening （HTS） due to no separation step, easy binding of drug targets and measuring their activities. Because of the diversification and commercialization of radio-labeled molecules and affinity tags, as well as the development of scintillation readers and liquid handling technologies, SPAs have been widely applied in receptor binding, high throughput drug screening, enzyme assay, radioimmunoassay, protein-protein interaction,cell-based assay and so on. This review presents the principle of SPA, discusses the key technologies（includes scintillation carrier, scintillation reader and radiolabel molecule） and analyzes the evaluation system, as well as outlines the assay development and provides some examples in details related to HTS, summarizes the shortcomings and gives some potential outlooks for the future study. Recently, SPA and fluorescence-based screenings have been the hot researches for the high throughput screening drug. The innovations on screening techniques will definitely promote our comprehensive understanding of cellular system biology and dramatically advance the discovery process of leading compounds.