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Updated: Jan 10, 2026

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays

Published on: October 23, 2019

Cellular Selectivity Analyses Reveal Distinguishing Profiles for Type II Kinase Inhibitors.

Matthew J Binder¹, Frances M Bashore¹, Kaitlin K Dunn Hoffman²

¹Structural Genomics Consortium, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

Biorxiv : the Preprint Server for Biology

|November 24, 2025

Summary

Assessing kinase inhibitor selectivity using cell-based assays provides different results than cell-free assays. This impacts chemical probe prioritization and reveals novel cellular kinase interactions.

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Area of Science:

Biochemistry
Pharmacology
Chemical Biology

Background:

Kinase inhibitor development requires rigorous selectivity assessment.
Historically, cell-free assays have been standard for kinase selectivity profiling.
There is a need to understand how cellular context affects inhibitor selectivity.

Purpose of the Study:

To compare kinase inhibitor selectivity profiling in cell-free versus cell-based assays.
To evaluate the impact of assay format on chemical probe prioritization.
To identify kinase interactions specific to the cellular environment.

Main Methods:

Profiling kinase inhibitors using traditional cell-free assays.
Utilizing a panel of cellular target engagement NanoBRET assays in intact cells.
Comparing selectivity data generated from both assay types.

Main Results:

Significant divergence observed between cell-free and cell-based selectivity profiling data.
Cellular assays influenced chemical probe prioritization compared to cell-free data.
Unanticipated kinase interactions were identified in cells for type II inhibitors, not seen in cell-free systems.

Conclusions:

Cellular target engagement assays provide a more relevant assessment of kinase inhibitor selectivity in a physiological context.
Switching from cell-free to cell-based assays can alter the landscape of identified on- and off-targets.
Cellular assays are crucial for uncovering context-specific drug-target interactions and improving chemical probe development.