Targeted Therapeutic Drug Discovery and Development Program, Division of Chemical Biology and Medicinal Chemistry, College of Pharmacy, The University of Texas at Austin, Austin, TX, USA.
SLAS Discov. 2021 Sep;26(8):984-994. doi: 10.1177/24725552211036056. Epub 2021 Jul 30.
Luminescence is characterized by the spontaneous emission of light resulting from either chemical or biological reactions. Because of their high sensitivity, reduced background interference, and applicability to numerous situations, luminescence-based assay strategies play an essential role in early-stage drug discovery. Newer developments in luminescence-based technologies have dramatically affected the ability of researchers to investigate molecular binding events. At the forefront of these developments are the nano bioluminescence resonance energy transfer (NanoBRET) and amplified luminescent proximity homogeneous assay (Alpha) technologies. These technologies have opened up numerous possibilities for analyzing the molecular biophysical properties of complexes in environments such as cell lysates. Moreover, NanoBRET enables the validation and quantitation of the interactions between therapeutic targets and small molecules in live cells, representing an essential benchmark for preclinical drug discovery. Both techniques involve proximity-based luminescence energy transfer, in which excited-state energy is transferred from a donor to an acceptor, where the efficiency of transfer depends on proximity. Both approaches can be applied to high-throughput compound screening in biological samples, with the NanoBRET assay providing opportunities for live-cell screening. Representative applications of both technologies for assessing physical interactions and associated challenges are discussed.
发光的特点是自发发射光，这是由化学或生物反应引起的。由于其灵敏度高、背景干扰小、适用于多种情况，基于发光的分析策略在药物早期发现中起着至关重要的作用。基于发光的技术的最新发展极大地影响了研究人员研究分子结合事件的能力。这些发展的前沿是纳米生物发光共振能量转移（NanoBRET）和放大发光邻近均相分析（Alpha）技术。这些技术为分析细胞裂解物等环境中复合物的分子生物物理特性开辟了许多可能性。此外，NanoBRET 能够验证和定量治疗靶标与小分子在活细胞中的相互作用，这是临床前药物发现的重要基准。这两种技术都涉及基于接近的发光能量转移，其中激发态能量从供体转移到受体，转移效率取决于接近度。这两种方法都可以应用于生物样品中的高通量化合物筛选，NanoBRET 分析提供了活细胞筛选的机会。讨论了这两种技术在评估物理相互作用和相关挑战方面的代表性应用。
