Generation of an antibody toolbox to characterize hERG

Hausammann, Georg J.; Heitkamp, Thomas; Matile, Hugues; Gsell, Bernard; Thoma, Ralf; Schmid, Georg; Frasson, David; Sievers, Martin; Hennig, Michael; Grütter, Markus G.

doi:10.1016/j.bbrc.2012.12.089

Publication type:	Article in scientific journal
Type of review:	Peer review (publication)
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.
DOI:	10.1016/j.bbrc.2012.12.089
Published in:	Biochemical and Biophysical Research Communications
Volume(Issue):	431
Issue:	1
Page(s):	70
Pages to:	75
Issue Date:	2013
Publisher / Ed. Institution:	Elsevier
ISSN:	0006-291X 1090-2104
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.
URI:	https://digitalcollection.zhaw.ch/handle/11475/11991
Fulltext version:	Published version
License (according to publishing contract):	Licence according to publishing contract
Departement:	Life Sciences and Facility Management
Organisational Unit:	Institute of Chemistry and Biotechnology (ICBT)
Appears in collections:	Publikationen Life Sciences und Facility Management

Files in This Item:

There are no files associated with this item.

Show full item record

Hausammann, G. J., Heitkamp, T., Matile, H., Gsell, B., Thoma, R., Schmid, G., Frasson, D., Sievers, M., Hennig, M., & Grütter, M. G. (2013). Generation of an antibody toolbox to characterize hERG. Biochemical and Biophysical Research Communications, 431(1), 70–75. https://doi.org/10.1016/j.bbrc.2012.12.089

Hausammann, G.J. et al. (2013) ‘Generation of an antibody toolbox to characterize hERG’, Biochemical and Biophysical Research Communications, 431(1), pp. 70–75. Available at: https://doi.org/10.1016/j.bbrc.2012.12.089.

G. J. Hausammann et al., “Generation of an antibody toolbox to characterize hERG,” Biochemical and Biophysical Research Communications, vol. 431, no. 1, pp. 70–75, 2013, doi: 10.1016/j.bbrc.2012.12.089.

HAUSAMMANN, Georg J., Thomas HEITKAMP, Hugues MATILE, Bernard GSELL, Ralf THOMA, Georg SCHMID, David FRASSON, Martin SIEVERS, Michael HENNIG und Markus G. GRÜTTER, 2013. Generation of an antibody toolbox to characterize hERG. Biochemical and Biophysical Research Communications. 2013. Bd. 431, Nr. 1, S. 70–75. DOI 10.1016/j.bbrc.2012.12.089

Hausammann, Georg J., Thomas Heitkamp, Hugues Matile, Bernard Gsell, Ralf Thoma, Georg Schmid, David Frasson, Martin Sievers, Michael Hennig, and Markus G. Grütter. 2013. “Generation of an Antibody Toolbox to Characterize hERG.” Biochemical and Biophysical Research Communications 431 (1): 70–75. https://doi.org/10.1016/j.bbrc.2012.12.089.

Hausammann, Georg J., et al. “Generation of an Antibody Toolbox to Characterize hERG.” Biochemical and Biophysical Research Communications, vol. 431, no. 1, 2013, pp. 70–75, https://doi.org/10.1016/j.bbrc.2012.12.089.