Development of a new and simple method for the detection of histidine-tagged proteins

Darain, Farzana, Ban, Changill and Shim, Yoon-Bo (2004) Development of a new and simple method for the detection of histidine-tagged proteins. Biosensors and Bioelectronics, 20 4: 856-862. doi:10.1016/j.bios.2004.03.028


Author Darain, Farzana
Ban, Changill
Shim, Yoon-Bo
Title Development of a new and simple method for the detection of histidine-tagged proteins
Journal name Biosensors and Bioelectronics   Check publisher's open access policy
ISSN 0956-5663
1873-4235
Publication date 2004-11-01
Sub-type Article (original research)
DOI 10.1016/j.bios.2004.03.028
Open Access Status Not yet assessed
Volume 20
Issue 4
Start page 856
End page 862
Total pages 7
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Language eng
Subject 1305 Biotechnology
1304 Biophysics
2204 Biomedical Engineering
1603 Electrochemistry
Abstract To develop a general method for the detection of histidine-tagged proteins, the interactions of the histidine epitope tag of MutH and MutL proteins with the epitope specific monoclonal anti-His antibody were monitored by a label-free direct method using impedance spectroscopy. The immunosensor was fabricated by covalent coupling of the antibody on a conducting polymer coated electrode surface. The impedance of the antibody modified electrode was decreased after binding to the histidine-tagged proteins. The specificity of the sensor was demonstrated by showing that no impedance change was occurred when the sensor was exposed to both of non-tagged MutH and MutL proteins. The specific interaction was further characterized using quartz crystal microbalance studies. Based on impedance measurements, the linear ranges were obtained from 50.0 to 125.0 and 50.0 to 250.0 μg/ml, for His-tag MutH and His-tag MutL proteins, respectively. The detection limits were determined to be 37.8 and 59.1 μg/ml, for His-tag MutH and His-tag MutL proteins, respectively.
Formatted abstract
To develop a general method for the detection of histidine-tagged proteins, the interactions of the histidine epitope tag of MutH and MutL proteins with the epitope specific monoclonal anti-His6 antibody were monitored by a label-free direct method using impedance spectroscopy. The immunosensor was fabricated by covalent coupling of the antibody on a conducting polymer coated electrode surface. The impedance of the antibody modified electrode was decreased after binding to the histidine-tagged proteins. The specificity of the sensor was demonstrated by showing that no impedance change was occurred when the sensor was exposed to both of non-tagged MutH and MutL proteins. The specific interaction was further characterized using quartz crystal microbalance studies. Based on impedance measurements, the linear ranges were obtained from 50.0 to 125.0 and 50.0 to 250.0 μg/ml, for His-tag MutH and His-tag MutL proteins, respectively. The detection limits were determined to be 37.8 and 59.1 μg/ml, for His-tag MutH and His-tag MutL proteins, respectively.
© 2004 Elsevier B.V. All rights reserved.
Keyword Histidine-tagged proteins
Impedance
Conducting polymer
Immunosensor
QCM
Label-free detection
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Australian Institute for Bioengineering and Nanotechnology Publications
 
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Created: Tue, 08 Feb 2011, 02:23:41 EST by Dr Farzana Darain on behalf of Aust Institute for Bioengineering & Nanotechnology