Hit Discovery, Discovery Sciences, R&D BioPharmaceuticals, AstraZeneca, Alderley Park, UK.
AstraZeneca R&D Discovery Sciences, Cambridge, UK.
Methods Mol Biol. 2021;2263:231-246. doi: 10.1007/978-1-0716-1197-5_10.
High-throughput assays based on fluorescence polarization (or fluorescence anisotropy) technology have often been employed for primary hit-finding in drug discovery. These binding assays provide a homogeneous format and consistent performance and offer advantages over some other optical methods. Developments in assay design and improvements in fluorescent probes have enabled the application of the technique to even complex biological systems. Here we describe the practical considerations for development of FP assays applied in high-throughput screening, including fluorophore selection, assay design, data analysis, and approaches for detecting compound interference.
基于荧光偏振（或荧光各向异性）技术的高通量检测方法常用于药物发现中的初步命中筛选。这些结合检测方法提供了均相格式和一致的性能，并优于其他一些光学方法。检测设计的发展和荧光探针的改进使该技术甚至能够应用于复杂的生物系统。在这里，我们将描述应用于高通量筛选的 FP 检测方法的开发的实际考虑因素，包括荧光团的选择、检测设计、数据分析以及检测化合物干扰的方法。
