Development of a Homogeneous High-Throughput Live-Cell G-Protein-Coupled Receptor Binding Assay
| Autor: | Carlo van Staden, Paul H. Lee, Steven C. Miller, Evan F. Cromwell |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Atropine
CHO Cells Muscarinic Antagonists Biology Ligands Biochemistry Receptors G-Protein-Coupled Analytical Chemistry chemistry.chemical_compound Cricetulus Cricetinae Muscarinic acetylcholine receptor Animals Receptor Fluorescent Dyes G protein-coupled receptor Miniaturization Drug discovery Ligand binding assay Chinese hamster ovary cell Benzenesulfonates Receptor Muscarinic M1 Parasympatholytics Pirenzepine Carbocyanines Ligand (biochemistry) Quinuclidinyl Benzilate chemistry Telenzepine Molecular Medicine Biological Assay Biotechnology |
| Zdroj: | SLAS Discovery. 13:748-754 |
| ISSN: | 2472-5552 |
| Popis: | The measurement of ligand receptor binding parameters for G-protein-coupled receptors is indispensable in the drug discovery process. Traditional ligand receptor binding assays require scale-up of cells and membrane preparations, which is an expensive and time-consuming process. In this report, the authors describe the development of a homogeneous live-cell binding assay for GPCRs using a fluorophore-labeled nonpeptide ligand. The model assay used Cy3B-labeled telenzepine and Chinese hamster ovary cells expressing M1 muscarinic acetylcholine receptors. This homogeneous live-cell fluorescence binding assay format is superior to the traditional binding methods because it measures binding of a ligand to intact receptors on living cells. The assay requires no washing or separation steps, thereby allowing a real-time kinetic readout for the determination of ligand association and dissociation from the intact receptors. The results also suggest that miniaturization is feasible without compromising the data quality. |
| Databáze: | OpenAIRE |
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