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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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.
|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|
|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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