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BRET technology to assess target engagement in live cells

Using Promega technology, scientists directly assess the dynamic process of target engagement in live cancer cells

A published paper using Promega NanoLuc® technology describes the first method to directly assess the dynamic process of target engagement in live cancer cells. As published in Nature Communications, Promega Scientists have developed a NanoLuc-based method for use in drug discovery that measures target residence time as a key aspect of target engagement in live cells. The technology uses bioluminescence resonance energy transfer (BRET), and allows measurement of drug binding to protein targets in real time and within live cells.

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NanoBRET® target engagement assay principle applied to intra-cellular targets. This image is used under the Creative Commons Attribution 4.0 International License. The image appears here in its original form as published in: Robers, M. B. et al. Target engagement and drug residence time can be observed in living cells with BRET. Nat. Commun. 6:10091 doi: 10.1038/ncomms10091 (2015).

To detect drug-target interactions, NanoBRET target engagement uses cell-permeable fluorescent tracers designed to interact with NanoLuc-tagged target proteins. The result of interaction is energy transfer from the NanoLuc protein to the tracer, generating a measurable signal. If a small molecule drug candidate interacts with the target protein, the tracer and drug compete for binding and BRET signal diminishes. Residence time measurement relies on pre-equilibration of compound with cells, removal of excess compound, and addition of tracer. Real-time signal monitoring shows compounds with slow target dissociation impede tracer binding and slow BRET signal production.

An additional publication in Nature Methods—"Application of BRET to monitor ligand binding to GPCRs"—details the application of NanoBRET technology. This article describes fluorescent ligand binding to NanoLuc® tagged cell-surface G-protein coupled receptors (GPCRs), an important drug target class.

A more recent paper in Nature Chemical Biology describes the use of NanoBRET target engagement approach and a novel BRET-based biosensor construct to elucidate the in cis bivalent binding mode of a class of bromodomain probe compounds that are capable of simultaneously binding to tandem bromodomains.

For more information on the NanoBRET Target Engagement Technology, visit





























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