| Sign In to gain access to subscriptions and/or personal tools. |
A Simple Theoretical Model for Fluorescence Polarization Binding Assay Development
Institut de Recherches Servier, Croissy-sur-Seine, France.
* To whom correspondence should be addressed. E-mail: olivier.nosjean{at}fr.netgrs.com.
Fluorescence polarization is a screening technology that is radioactivity free, homogeneous, and ratiometric. The signal measured with this technology is a weighted value of free and bound ligand. As a consequence, saturation curves are accessible only after calculation of the corresponding concentrations of free and bound ligand. To make this technology more accessible to assay development, the authors propose a simple mathematical model that predicts fluorescence polarization values from ligand and receptor total concentrations, depending on the corresponding dissociation constant. This model was validated using data of Bodipy-NDP- Key Words: fluorescence polarization, binding assay, assay development, mathematical model, MC5
First published on November 7, 2006, doi:10.1177/1087057106294841 |
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
|
|
 |
|
Sign In to gain access to subscriptions and/or personal tools.
MSH binding to MC5, obtained after either ligand saturation of a receptor preparation or, conversely, receptor saturation of a ligand solution. These experimental data were also used to calculate the actual concentration of free and bound ligand and receptor and to obtain pharmacological constants by Scatchard analysis. A general method is proposed, which facilitates the design of fluorescence polarization binding assays by relying on the representation of theoretical polarization values. This approach is illustrated by the application to 2 systems of very different affinities.