IMR Press / FBS / Volume 8 / Issue 2 / DOI: 10.2741/S462

Frontiers in Bioscience-Scholar (FBS) is published by IMR Press from Volume 13 Issue 1 (2021). Previous articles were published by another publisher on a subscription basis, and they are hosted by IMR Press on as a courtesy and upon agreement with Frontiers in Bioscience.


Moderate to high throughput in vitro binding kinetics for drug discovery

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1 In vitro Pharmacology, Merck Research Laboratories, 2015 Galloping Hill Road, Kenilworth, New Jersey, USA

*Author to whom correspondence should be addressed.


Front. Biosci. (Schol Ed) 2016, 8(2), 278–297;
Published: 1 June 2016

This review provides a concise summary for state of the art, moderate to high throughput in vitro technologies being employed to studydrug-target binding kinetics. These technologies cover a wide kinetic timescale spanning up to nine orders of magnitude from milliseconds to days. Automated stopped flow measurestransient and (pre)steady state kinetics from milliseconds to seconds. For seconds to hours timescale kinetics we discuss surface plasmon resonance-based biosensor,global progress curve analysis for high throughput kinetic profiling of enzyme inhibitors and activators, and filtration plate-based radioligand or fluorescentbinding assays for receptor binding kinetics. Jump dilution after pre-incubation is the preferred method for very slow kinetics lasting for days. The basic principles,best practices and simulated data for these technologies are described. Finally, the application of a universal label-free technology, liquid chromatography coupledtandem mass spectrometry (LC/MS/MS), is briefly reviewed. Select literature references are highlighted for in-depth understanding. A new reality is dawning whereinbinding kinetics is an integral and routine part of mechanism of action elucidation and translational, quantitative pharmacology for drug discovery.

Binding Kinetics
Stopped Flow
Surface Plasma Resonance
Global Progress Curve Analysis
Radioligand Binding
Jump Dilution
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