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

被引：28

作者：

Darain, F

Ban, CI ^{[1
]}

Shim, YB

机构：

[1] Pusan Natl Univ, Dept Chem, Pusan 609735, South Korea

[2] Pusan Natl Univ, Ctr Innovat Biophys Sensor Technol, Pusan 609735, South Korea

[3] Pohang Univ Sci & Technol, Dept Chem, Pohang, South Korea

[4] Pohang Univ Sci & Technol, Div Mol & Life Sci BK21, Pohang, South Korea

来源：

BIOSENSORS & BIOELECTRONICS | 2004年 / 20卷 / 04期

关键词：

histidine-tagged proteins; impedance; conducting polymer; immunosensor; QCM; label-free detection;

D O I：

10.1016/j.bios.2004.03.028

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

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(6) antibody were monitored by a label-free direct method using impedance spectroscopy. The immumosensor 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 mug/ml, for His-tag MutH and His-tag MutL proteins, respectively. The detection limits were determined to be 37.8 and 59.1 mug/ml, for His-tag MutH and His-tag MutL proteins, respectively. (C) 2004 Elsevier B.V. All rights reserved.

引用

页码：857 / 863

页数：7

共 18 条

[1] Structural basis for MutH activation in E-coli mismatch repair and relationship of MutH to restriction endonucleases [J].