Current in vitro kinase assay technologies: the quest for a universal format.
Current in vitro kinase assay technologies: the quest for a universal format.
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DOI：
10.2174/157016308783769414
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发表时间：
2008-03-01
影响因子：
--
通讯作者：
Talanian, Robert V
中科院分区：
文献类型：
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作者：
Jia, Yong;Quinn, Christopher M;Talanian, Robert V
对激酶作为药物靶点的兴趣迅速增长，促进了许多激酶测定技术的发展。这些技术可分为三类：放射性测定、磷酸化抗体依赖性荧光/发光测定和磷酸化抗体非依赖性荧光/发光测定。本文将回顾市场上一些主要的激酶测定技术，特别强调最新的系统。我们将描述这些技术的物理原理，实际的优点和缺点，以及这些技术在激酶药物发现中的潜在应用。这些技术大多适用于高温超导，但只有少数可以用于动力学和机理研究。近年来，在开发不含放射性同位素和定制试剂（如磷酸特异性抗体）的通用测定方面取得了重大进展。然而，由于每种形式的各种限制，这些通用测定技术中没有一种可以声称是真正通用的。在采用特定的激酶检测技术之前，应考虑几个因素，包括预期的应用、成本、时间轴、专业知识、熟悉程度和舒适度。
The rapidly growing interest in kinases as drug targets has prompted the development of many kinase assay technologies. These technologies can be grouped into three categories: radiometric assays, phospho-antibody-dependent fluorescence/luminescence assays, and phospho-antibody-independent fluorescence/luminescence assays. This article will review some of the major kinase assay technologies on the market, with particular emphasis on the newest systems. We will describe the physical principles, the practical advantages and drawbacks, and the potential applications of these technologies in kinase drug discovery. Most of these technologies are suitable for HTS, but only a few can be utilized for kinetic and mechanistic studies. Significant progress towards development of generic assays, free of radioisotopes and custom reagents such as phospho-specific antibodies, has been made in recent years. However, due to various limitations of each format, none of these generic assay technologies can yet claim to be truly universal. Several factors, including the intended applications, cost, timeline, expertise, familiarity, and comfort level, should be considered prior to pursuing a particular kinase assay technology.