Current advances in the use of bioluminescence assays for drug discovery: an update of the last ten years

ABSTRACTIntroduction 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.Areas covered 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.Expert opinion 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.KEYWORDS: BioluminescenceBRET assayscell-free assaysdrug screeningmultiplexingprotein-complementation assaysreporter gene assay3D cell models Article highlightsBioluminescent assays are a powerful tool for drug screening assaysA wide portfolio of luciferases is now available with tuned emission wavelengths, kinetics and high stabilityBioluminescent cell-free and cell-based assays can be implemented in the early stages of the drug discovery processSplit bioluminescence complementation assays and resonance energy transfer assays represent viable approaches for monitoring protein-protein interactionsBioluminescence multiplexing allows to simultaneously monitor multiple targets and mechanisms of actionsThe implementation of bioluminescence in 3D cell models provides a new level of predictivity in drug discovery programmesDeclaration of interestThe authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.Reviewer disclosuresPeer reviewers on this manuscript have no relevant financial or other relationships to disclose.Additional informationFundingThis manuscript has not been funded.