Determination of dissociation constants for protein-ligand complexes by electrospray ionization mass spectrometry

Tjernberg, A., Carno, S., Oliv, F., Benkestock, K., Edlund, P-O., Griffiths, W.J. and Hallen, D. (2004) Determination of dissociation constants for protein-ligand complexes by electrospray ionization mass spectrometry. Analytical Chemistry, 76 . pp. 4325-4331. 10.1021/ac0497914.

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DOI: 10.1021/ac0497914

Abstract

Determination of Dissociation Constants for Protein-Ligand Complexes by Electrospray Ionization Mass Spectrometry Agneta Tjernberg,* Sofi Carnö, Frida Oliv, Kurt Benkestock, Per-Olof Edlund, William J. Griffiths, and Dan Hallén Departments of Structural Chemistry and Preclinical R&D, Biovitrum AB, SE-112 76 Stockholm, Sweden Received for review February 6, 2004. Accepted May 11, 2004. Abstract: A fully automated biophysical assay based on electrospray ionization mass spectrometry (ESI-MS) for the determination of the dissociation constants (KD) between soluble proteins and low molecular mass ligands is presented. The method can be applied to systems where the relative MS response of the protein and the protein-ligand complexes do not reflect relative concentrations. Thus, the employed approach enables the use of both electrostatically and nonpolar bound complexes. The dynamic range is wider than for most biological assays, which facilitates the process of establishing a structure-activity relationship. This fully automated ESI-MS assay is now routinely used for ligand screening. The entire procedure is described in detail using hGHbp, a 25-kDa extracellular soluble domain of the human growth hormone receptor, as a model protein.

Item Type:Article
Additional Information:Full text available in print and electronically at the School of Pharmacy Libray.
Departments, units and centres:Department of Pharmaceutical and Biological Chemistry > Department of Pharmaceutical and Biological Chemistry
ID Code:636
Journal or Publication Title:Analytical Chemistry
Deposited By:Library Staff
Deposited On:19 Jan 2007
Last Modified:19 Sep 2007 11:15

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