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Related Experiment Video

Updated: Jul 19, 2025

Identification of Kinase-substrate Pairs Using High Throughput Screening
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NanoBRET™ Live-Cell Kinase Selectivity Profiling Adapted for High-Throughput Screening.

Amanda N Nieman1, Kaitlin K Dunn Hoffman1, Elizabeth R Dominguez1

  • 1Promega Corporation, Madison, WI, USA.

Methods in Molecular Biology (Clifton, N.J.)
|August 9, 2023
PubMed
Summary

This study enhances live-cell kinase selectivity profiling using automation and acoustic dispensing. The scalable workflow significantly increases throughput for drug discovery efforts targeting kinases.

Keywords:
BRETHigh-Throughput Screening (HTS)Kinase profilingLive-cell target engagementNanoBRETTM

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Area of Science:

  • Biochemistry
  • Pharmacology
  • Drug Discovery

Background:

  • Kinases are key targets in drug discovery, but achieving intracellular selectivity for inhibitors is challenging.
  • The NanoBRET platform offers live-cell kinase selectivity profiling but is limited by manual workflows.
  • High-throughput screening is crucial for identifying effective kinase inhibitors.

Purpose of the Study:

  • To develop a scalable and automated workflow for live-cell kinase selectivity profiling.
  • To enhance the throughput of the NanoBRET platform for broader compound screening.
  • To enable robust quantitative analysis of kinase inhibitor selectivity.

Main Methods:

  • Implementation of automation and acoustic dispensing for liquid handling.
  • Adaptation of the NanoBRET platform for high-throughput screening.
  • Profiling of compound sets against 192 full-length protein kinases in live cells.

Main Results:

  • Dramatically improved throughput compared to manual liquid handling.
  • Enabled profiling of larger compound libraries against a wide range of kinases.
  • Achieved statistical robustness supporting quantitative kinase selectivity analysis.

Conclusions:

  • The automated NanoBRET workflow significantly enhances kinase inhibitor screening capabilities.
  • This scalable approach facilitates more comprehensive drug discovery efforts targeting kinases.
  • The optimized workflow supports efficient identification of selective kinase inhibitors.