First Assay to Quantitatively Measure Drug-Kinase Interactions in Live Cells

Promega Corporation’s NanoBRET™ Target Engagement (TE) Intracellular Kinase Assay is the first assay to quantify inhibitor drug binding to kinase proteins inside live human cells, as demonstrated in the recent Cell Chemical Biology article, “Quantitative, Wide-spectrum Kinase Profiling in Live Cells for Assessing the Effect of Cellular ATP on Target Engagement.” In collaboration with Structural Genomics Consortium (SGC), this article shows the greater predictive potential of NanoBRET Target Engagement compared to common traditional approaches that use purified kinase domains in non-cell-based assays.

“The NanoBRET TE assay is a major advance in our ability to demonstrate the potency and selectivity of kinase inhibitors in living cells,” says Tim Willson, Chief Scientist SGC-UNC. “Application of this technology to development of chemical probes will accelerate our study of the hundreds of dark kinases that may be new targets for drug discovery.”

NanoBRET TE uses bioluminescence resonance energy transfer (BRET) and allows measurement of drug binding to protein targets in real time inside live cells using a simplified experimental protocol. Accurate measurement of drug-target engagement in live cells is critical to understanding the biology of drug interactions against target proteins, with the ultimate aim of developing better therapies for human diseases.

The approach has proven beneficial for researchers at SGC-Frankfurt who are currently testing broad-spectrum profiling of specific kinase inhibitors in live cells for a more realistic estimate of inhibitor selectivity under more physiological conditions.

“We have already gained valuable insights regarding the kinetics of specific kinase inhibitors, and we also see great potential for this assay for other target families,” says Susanne Müller-Knapp, Senior Project Manager Chemical Probes, SGC Frankfurt.

The NanoBRET TE platform combines the sensitivity of NanoLuc^® technology with quantitative capabilities of energy transfer. Unlike other methods, NanoBRET TE Kinase Assay allows for quantitative measurements of drug affinity and kinase selectivity inside live cells at target levels similar to that of endogenous kinases. To support kinase biology, Promega has just launched more than 100 individual full-length kinase target engagement assays.

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