Interference of metal ions on the bioluminescent signal of firefly, Renilla, and NanoLuc luciferases in high-throughput screening assays.

ABSTRACT: Bioluminescent high-throughput screening (HTS) assays, based largely on the activity of firefly (FLuc), Renilla (RLuc), and/or NanoLuc (NLuc) luciferases, are widely utilised in research and drug discovery. In this study, we quantify the luciferase-based real-life HTS assay interference from biologically and environmentally relevant metal ions ubiquitously present in buffers, environmental and biological matrices, and as contaminants in plastics and compound libraries. We also provide insights into the cross-effects of metal ions and other key experimental and biological reagents (e.g., buffer types, EDTA, and glutathione) to inform HTS assay design, validation, and data interpretation. A total of 21 ions were screened in three robust HTS assays ("SC" assays) based on the luminescence of F

SUBMITTER: Canyelles I Font F 

PROVIDER: S-EPMC11628255 | biostudies-literature | 2024

REPOSITORIES: biostudies-literature