Title: Generation of an antibody toolbox to characterize hERG
Authors : Hausammann, Georg J.
Heitkamp, Thomas
Matile, Hugues
Gsell, Bernard
Thoma, Ralf
Schmid, Georg
Frasson, David
Sievers, Martin
Hennig, Michael
Grütter, Markus G.
Published in : Biochemical and biophysical research communications
Volume(Issue) : 431
Issue : 1
Pages : 70
Pages to: 75
Publisher / Ed. Institution : Elsevier
Issue Date: 2013
License (according to publishing contract) : Licence according to publishing contract
Type of review: Peer review (Publication)
Language : English
Subjects : Monoclonal antibodies; Gel chromatography; ERG1 potassium channel; Ether-A-Go-Go potassium channels; HEK293 cells; Enzyme-linked immunosorbent assay
Subject (DDC) : 571: Physiology and related subjects
Abstract: The human ether-a-go-go related gene (hERG) potassium channel plays a major role in the repolarization of the cardiac action potential. Inhibition of the hERG function by mutations or a wide variety of pharmaceutical compounds cause long QT syndrome and lead to potentially lethal arrhythmias. For detailed insights into the structural and biochemical background of hERG function and drug binding, the purification of recombinant protein is essential. Because the hERG channel is a challenging protein to purify, fast and easy techniques to evaluate different expression, solubilization and purification conditions are of primary importance. Here, we describe the generation of a set of 12 monoclonal antibodies against hERG. Beside their suitability in western blot, immunoprecipitation and immunostaining, these antibodies were used to establish a sandwich ELISA for the detection and relative quantification of hERG in different expression systems. Furthermore, a Fab fragment was used in fluorescence size exclusion chromatography to determine the oligomeric state of hERG after solubilization. These new tools can be used for a fast and efficient screening of expression, solubilization and purification conditions.
Departement: Life Sciences and Facility Management
Organisational Unit: Institute of Chemistry and Biotechnology (ICBT)
Publication type: Article in scientific Journal
DOI : 10.1016/j.bbrc.2012.12.089
ISSN: 0006-291X
1090-2104
URI: https://digitalcollection.zhaw.ch/handle/11475/11991
Appears in Collections:Publikationen Life Sciences und Facility Management

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