Hall Matthew D, Simeonov Anton, Davis Mindy I
NCATS Chemical Genomics Center, National Center for Advancing Translational Sciences, National Institutes of Health , Rockville, Maryland.
Assay Drug Dev Technol. 2016 Apr;14(3):175-9. doi: 10.1089/adt.2016.707.
Fluorescence is utilized as the output for a range of assay formats used in high-throughput screening (HTS). Interference with these assays from the compounds in libraries utilized in HTS is a well-recognized phenomenon, particularly for assays relying on UV excitation such as for direct detection of the oxidoreductase cofactors NADH or NADPH. In this study, we discuss these interference challenges and highlight the specific case of the diaphorase/resazurin system that can be coupled to enzymes utilizing NADH or NADPH. We review the utilization of this assay system in the literature and argue that the diaphorase/resazurin system is underutilized in assay development. It is the authors' hope that this Perspective and the accompanying Technical Brief in this issue will stimulate interest in a robust and sensitive coupling system to avoid assay fluorescence interference.
荧光被用作高通量筛选（HTS）中一系列检测形式的输出信号。高通量筛选中使用的化合物库对这些检测产生干扰是一种公认的现象，特别是对于依赖紫外线激发的检测，例如直接检测氧化还原酶辅因子NADH或NADPH的检测。在本研究中，我们讨论了这些干扰挑战，并突出了可与利用NADH或NADPH的酶偶联的黄递酶/刃天青系统的具体情况。我们回顾了该检测系统在文献中的应用，并认为黄递酶/刃天青系统在检测开发中未得到充分利用。作者希望本期的这篇观点文章及随附的技术简报能激发人们对一种强大且灵敏的偶联系统的兴趣，以避免检测荧光干扰。
