Identification of drug-binding sites on human serum albumin using affinity capillary electrophoresis and chemically modified proteins as buffer additives

Electrophoresis
Hee Seung KimD S Hage

Abstract

A technique based on affinity capillary electrophoresis (ACE) and chemically modified proteins was used to screen the binding sites of various drugs on human serum albumin (HSA). This involved using HSA as a buffer additive, following the site-selective modification of this protein at two residues (tryptophan 214 or tyrosine 411) located in its major binding regions. The migration times of four compounds (warfarin, ibuprofen, suprofen and flurbiprofen) were measured in the presence of normal or modified HSA. These times were then compared and the mobility shifts observed with the modified proteins were used to identify the binding regions of each injected solute on HSA. Items considered in optimizing this assay included the concentration of protein placed into the running buffer, the reagents used to modify HSA, and the use of dextran as a secondary additive to adjust protein mobility. The results of this method showed good agreement with those of previous reports. The advantages and disadvantages of this approach are examined, as well as its possible extension to other solutes.

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Citations

Nov 25, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·C BertucciV Andrisano
Apr 15, 2004·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Jian-Feng ZhangYing-Tang Lu
Feb 28, 2014·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Xiwei ZhengDavid S Hage
Aug 28, 2003·Bioorganic Chemistry·M DelfiniG Rugo
Feb 5, 2003·Current Opinion in Biotechnology·Chiara Galbusera, David D Y Chen
Nov 25, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Shufeng Zhou
Jun 17, 2008·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Hee Seung Kim, Irving W Wainer
Jun 12, 2009·Electrophoresis·Niels H H Heegaard
Dec 7, 2006·Electrophoresis·Lotta K Amundsen, Heli Sirén
Feb 14, 2013·Chemistry, an Asian Journal·Dipanwita DeAnindya Datta
Jan 25, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Hee Seung Kim, David S Hage
May 12, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Hee Seung KimDavid S Hage
Jul 1, 2011·Analytical Methods : Advancing Methods and Applications·David S HageXiwei Zheng

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