■生物发光是一种成熟的光学检测技术,广泛用于多种生物分析应用中,包括高通量和高内容筛选。由于合成生物学技术和深度学习的进步,现在可以使用具有调谐发射波长的广泛的荧光素酶组合,动力学,和高稳定性。这些荧光素酶可以在药物发现和开发管道中实施,允许高灵敏度和复用能力。
■这篇综述总结了生物发光系统作为体外应用药物发现计划中的工具集的最新进展。特别关注过去10年(2013年至2023年)中用于药物筛选的最先进的基于生物发光的技术,例如无细胞测定,基于转基因细胞的基于细胞的检测,生物发光共振能量转移,以及2D和3D细胞模型中的蛋白质互补分析。
■具有改善的发射和稳定性特性的调谐生物发光蛋白的可用性对于开发用于药物发现的生物发光测定法至关重要,从报告基因技术到蛋白质-蛋白质技术。进一步研究,将机器学习与合成生物学相结合,将有必要获得可持续和高度预测性生物发光药物发现平台的新工具。
Bioluminescence is a well-established optical detection technique widely used in several bioanalytical applications, including high-throughput and high-content screenings. Thanks to advances in synthetic biology techniques and deep learning, a wide portfolio of luciferases is now available with tuned emission wavelengths, kinetics, and high stability. These luciferases can be implemented in the drug discovery and development pipeline, allowing high sensitivity and multiplexing capability.
This review summarizes the latest advancements of bioluminescent systems as toolsets in drug discovery programs for in vitro applications. Particular attention is paid to the most advanced bioluminescence-based technologies for drug screening over the past 10 years (from 2013 to 2023) such as cell-free assays, cell-based assays based on genetically modified cells, bioluminescence resonance energy transfer, and protein complementation assays in 2D and 3D cell models.
The availability of tuned bioluminescent proteins with improved emission and stability properties is vital for the development of bioluminescence assays for drug discovery, spanning from reporter gene technology to protein-protein techniques. Further studies, combining machine learning with synthetic biology, will be necessary to obtain new tools for sustainable and highly predictive bioluminescent drug discovery platforms.