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High-throughput screening with HyperCyt flow cytometry to detect small molecule formylpeptide receptor ligands
Susan M Young1, Cristian Bologa, Eric R Prossnitz
1Cytometry and Department of Pathology, Cancer Research and Treatment Center, University of New Mexico, Health Sciences Center, Albuquerque, NM 87131, USA.
Journal of Biomolecular Screening
|June 21, 2005
Summary
High-throughput flow cytometry (HTFC) using the HyperCyt system enables rapid screening of compound libraries. This robust method effectively identifies anti-inflammatory compounds targeting formylpeptide receptors (FPR).
Area of Science:
- Biochemistry
- Pharmacology
- Immunology
Background:
- The formylpeptide receptor (FPR) family mediates leukocyte recruitment in chronic inflammation.
- High-throughput flow cytometry (HTFC) offers a powerful approach for compound library screening.
- Automated systems are crucial for efficient analysis in HTFC.
Purpose of the Study:
- To develop and apply an HTFC screening method for identifying anti-inflammatory compounds.
- To detect compounds that inhibit ligand binding to formylpeptide receptors (FPR).
- To validate the efficiency and sensitivity of the HyperCyt system for library screening.
Main Methods:
- Development of a homogeneous no-wash assay for HTFC.
- Utilizing the HyperCyt automated system for rapid sample aspiration and processing.
- Screening of the Prestwick Chemical Library (880 compounds) for FPR antagonists.
Main Results:
- Routine sample processing at 1.5 seconds per well, analyzing approximately 2500 cells/sample.
- Complete processing of a 96-well plate in under 2.5 minutes.
- Successful identification of a known FPR antagonist (sulfinpyrazone) with low affinity (Ki=14 µM).
Conclusions:
- The HyperCyt system provides a robust, sensitive, and quantitative HTFC method.
- This approach is highly effective for screening compound libraries in a 96-well format.
- The developed assay is suitable for identifying novel anti-inflammatory agents targeting FPR.