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Published on: September 9, 2021
A Simple and robust automated kinase profiling platform using luminescent ADP accumulation technology.
Brad Larson1, Peter Banks, Hicham Zegzouti
1BioTek Instruments, Inc., Winooski, Vermont, USA. larsonb@biotek.com
Assay and Drug Development Technologies
|January 12, 2010
Summary
This study presents a versatile luminescent assay for kinase profiling, reducing optimization time and costs. The method uses a single reagent set for diverse kinases, enabling efficient drug discovery.
Area of Science:
- Biochemistry
- Drug Discovery
- Enzymology
Background:
- Kinases are crucial targets in drug discovery.
- Profiling lead compounds against various kinases is essential to identify off-target effects.
- Current kinase assays often require specific substrates, increasing optimization time and costs.
Purpose of the Study:
- To demonstrate a versatile luminescent ADP accumulation assay for kinase profiling.
- To showcase a method that reduces optimization time and costs.
- To present an automated solution for comprehensive kinase profiling.
Main Methods:
- Utilized a luminescent ADP accumulation assay adaptable to various kinases.
- Employed a single set of reagents for kinases with different ATP and substrate K(m app) values.
- Automated the assay process using a simple, inexpensive pipetting system.
Main Results:
- The assay demonstrated versatility across a wide range of kinases.
- High assay sensitivity allowed for low enzyme concentrations and minimal ATP conversion with good signal-to-background ratios.
- Automated profiling generated compound IC(50) values that agreed with literature data.
Conclusions:
- The luminescent ADP accumulation assay offers a robust and efficient solution for kinase profiling.
- This automated approach simplifies and accelerates the identification of potential off-target effects.
- The method provides a cost-effective and versatile tool for drug discovery efforts.
