Liu Ya-Li, Guo Zhan-Yun
Research Center for Translational Medicine at East Hospital, College of Life Sciences and Technology, Tongji University, Shanghai, China.
Methods Mol Biol. 2016;1461:65-90. doi: 10.1007/978-1-4939-3813-1_6.
Bioluminescence has been widely used in biomedical research due to its high sensitivity, low background, and broad linear range. In recent studies, we applied bioluminescence to ligand-receptor binding assays for some protein or peptide hormones based on a newly developed small monomeric Nanoluciferase (NanoLuc) reporter that has the so far brightest bioluminescence. The conventional ligand-receptor binding assays rely on radioligands that have drawbacks, such as radioactive hazards and short shelf lives. In contrast, the novel bioluminescent binding assays use the NanoLuc-based protein or peptide tracers that are safe, stable, and ultrasensitive. Thus, the novel bioluminescent ligand-receptor binding assay would be applied to more and more protein or peptide hormones for ligand-receptor interaction studies in future. In the present article, we provided detailed protocols for setting up the novel bioluminescent ligand-receptor binding assays using two representative protein hormones as examples.
由于生物发光具有高灵敏度、低背景和宽线性范围,它已被广泛应用于生物医学研究。在最近的研究中,我们基于一种新开发的小单体纳米荧光素酶(NanoLuc)报告基因,将生物发光应用于一些蛋白质或肽类激素的配体-受体结合测定,该报告基因具有迄今为止最亮的生物发光。传统的配体-受体结合测定依赖于放射性配体,而放射性配体存在诸如放射性危害和保质期短等缺点。相比之下,新型生物发光结合测定使用基于NanoLuc的蛋白质或肽类示踪剂,这些示踪剂安全、稳定且超灵敏。因此,新型生物发光配体-受体结合测定在未来将应用于越来越多的蛋白质或肽类激素,用于配体-受体相互作用研究。在本文中,我们以两种代表性蛋白质激素为例,提供了建立新型生物发光配体-受体结合测定的详细方案。