Chapter title |
Single Step Determination of Unlabeled Compound Kinetics Using a Competition Association Binding Method Employing Time-Resolved FRET
|
---|---|
Chapter number | 10 |
Book title |
Rational Drug Design
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-8630-9_10 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8629-3, 978-1-4939-8630-9
|
Authors |
David A. Sykes, Steven J. Charlton, Sykes, David A., Charlton, Steven J. |
Abstract |
The competition association binding method allows the characterization of the kinetics of unlabeled compounds and the calculation of receptor-drug affinity (K d). The K d value is defined as the ratio of the dissociation constant (or k off) of the receptor-bound ligand to its association rate constant (or k on) for a system at equilibrium. Traditionally, competition association binding experiments have been carried out using radiometric detection methods with limited assay throughput. Here we describe a novel method for the determination of unlabeled compound kinetics using the technique of time-resolved fluorescence resonance energy transfer (TR-FRET) performed at physiological temperature and sodium ion concentration. Based on a traditional screening format (10-point curves), up to 28 compounds can be tested on a single 384-well plate by this method. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 13 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 5 | 38% |
Student > Ph. D. Student | 4 | 31% |
Student > Bachelor | 1 | 8% |
Professor | 1 | 8% |
Student > Master | 1 | 8% |
Other | 1 | 8% |
Readers by discipline | Count | As % |
---|---|---|
Pharmacology, Toxicology and Pharmaceutical Science | 6 | 46% |
Biochemistry, Genetics and Molecular Biology | 3 | 23% |
Computer Science | 1 | 8% |
Neuroscience | 1 | 8% |
Unknown | 2 | 15% |