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Updated: Jun 27, 2026
Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
Published on: October 23, 2019
High-throughput biochemical kinase selectivity assays: panel development and screening applications
Amy Card1, Chris Caldwell, Hyunsuk Min
1Pfizer Research Technology Center, Cambridge, Massachusetts, USA.
Abstract:
Kinases represent attractive targets for drug discovery. Eight small-molecule kinase inhibitors are currently marketed in the area of oncology, and numerous others are in clinical trials. Characterization of the selectivity profiles of these compounds is important to target appropriate patient populations and to reduce the potential of toxicity due to off-target effects. The authors describe the development, validation, and utilization of a biochemical kinase assay panel for the selectivity profiling of inhibitors. The panel was developed as 29 radiometric Flashplate assays, and then an initial 13 were transitioned to a nonradiometric Caliper mobility shift assay format. Generation of high-quality data from the panel is detailed along with a comparison of the assay formats. Both assay technologies were found to be suitable for panel screening, but mobility shift assays yielded higher data quality. The selectivity data generated here should be useful in computational modeling and help facilitate, in conjunction with sequence and structural information, the rational design of inhibitors with well-defined selectivity profiles.
Insights
Researchers developed a biochemical kinase assay panel to profile small-molecule inhibitors. Mobility shift assays provided higher data quality than radiometric assays, aiding rational drug design for oncology kinase inhibitors.
Area of Science:
- Biochemistry
- Pharmacology
- Drug Discovery
Background:
- Kinase inhibitors are crucial in oncology drug discovery, with eight marketed and many in clinical trials.
- Understanding inhibitor selectivity is vital for effective patient targeting and minimizing off-target toxicity.
Purpose of the Study:
- To develop and validate a biochemical kinase assay panel for selectivity profiling of small-molecule inhibitors.
- To compare the performance of radiometric and non-radiometric assay formats for kinase inhibitor screening.
Main Methods:
- Development of a 29-assay radiometric Flashplate panel.
- Transition of 13 assays to a non-radiometric Caliper mobility shift assay format.
- Generation and quality assessment of selectivity data using both assay formats.
Main Results:
- Both radiometric Flashplate and Caliper mobility shift assays are suitable for kinase inhibitor panel screening.
- Mobility shift assays demonstrated superior data quality compared to radiometric assays.
- The generated selectivity data are valuable for computational modeling and rational inhibitor design.
Conclusions:
- The developed kinase assay panel provides high-quality selectivity data for small-molecule inhibitors.
- Mobility shift assays offer advantages in data quality for kinase inhibitor profiling.
- These findings support the rational design of kinase inhibitors with improved selectivity profiles for oncology applications.
