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Updated: Jul 8, 2026
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Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery
Published on: October 4, 2018
High-content single-cell drug screening with phosphospecific flow cytometry
Peter O Krutzik1, Janelle M Crane, Matthew R Clutter
1Department of Microbiology and Immunology, Baxter Laboratory in Genetic Pharmacology, Stanford University, 269 Campus Drive, Stanford, California 94305, USA.
Nature Chemical Biology
|December 25, 2007
Summary
This study introduces a cell-based drug discovery platform using phosphospecific flow cytometry (phosphoflow) to screen for kinase inhibitors. The platform identified selective inhibitors for Jak-Stat and MAP kinase pathways in primary cells and validated them in vivo.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- Traditional drug screening often uses simplified cell-free assays.
- Cell-based systems are crucial for understanding complex cellular networks and disease states.
- Endogenous kinase signaling pathways are key targets in drug discovery.
Purpose of the Study:
- To develop and validate a high-content, cell-based drug discovery platform.
- To screen for pathway-selective kinase inhibitors using phosphospecific flow cytometry (phosphoflow).
- To identify inhibitors targeting Jak-Stat and MAP kinase signaling pathways.
Main Methods:
- Utilized phosphospecific flow cytometry (phosphoflow) for high-content, single-cell analysis.
- Screened a library of small-molecule natural products against heterogeneous primary cell populations.
- Conducted dose-response experiments to assess pathway selectivity and cell-type differential inhibition.
- Performed in vivo validation of lead compound selectivity in mice.
Main Results:
- Identified pathway-selective inhibitors of Jak-Stat and MAP kinase signaling.
- Demonstrated differential inhibition of various cell types by identified compounds.
- Revealed novel druggability trends across multiple small molecules.
- Confirmed lead compound selectivity in vivo.
Conclusions:
- Phosphoflow is a versatile platform for cell-based drug discovery.
- The platform enables screening, validation, and monitoring of drug efficacy throughout the discovery process.
- This approach advances the study of kinase signaling and identifies potential therapeutic agents.
