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Sensors 2012, 12(8), 10097-10108; doi:10.3390/s120810097

Label-Free Electrochemical Diagnosis of Viral Antigens with Genetically Engineered Fusion Protein

2,*  and 7,*
1 BioProcess Engineering Research Center, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Korea 2 Department of Chemical & Biological Engineering, Chungnam National University, 99 Daehangno, Yuseong-gu, Daejeon 305-764, Korea 3 Center for Nanobio Integration & Convergence Engineering, National Nanofab Center, 291 Daehak-ro, Yuseong-gu, Daejeon 305-806, Korea 4 Department of Chemical & Biomolecular Engineering (BK21 Program), Department of Bio & Brain Engineering, Department of Biological Sciences, Bioinformatics Research Center, Center for Systems & Synthetic Biotechnology, and Institute for the BioCentury, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Korea 5 Department of Microbiology & Research Institute for Medical Science, Chungnam National University, 6 Moonhwa-dong, Jung-gu, Daejeon 301-747, Korea 6 Biotechnology Research Division, National Fisheries Research & Development Institute (NFRDI), 408-1 Sirang-ri, Gijang, Busan 619-705, Korea 7 Department of Chemistry, Chung-Ang University, 84 Heukseok-ro, Dongjak-gu, Seoul 156-756, Korea Theses authors contributed equally to this work.
* Authors to whom correspondence should be addressed.
Received: 10 May 2012 / Revised: 12 July 2012 / Accepted: 15 July 2012 / Published: 26 July 2012
(This article belongs to the Section Biosensors)
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We have developed a simple electrochemical biosensing strategy for the label-free diagnosis of hepatitis B virus (HBV) on a gold electrode surface. Gold-binding polypeptide (GBP) fused with single-chain antibody (ScFv) against HBV surface antigen (HBsAg), in forms of genetically engineered protein, was utilized. This GBP-ScFv fusion protein can directly bind onto the gold substrate with the strong binding affinity between the GBP and the gold surface, while the recognition site orients toward the sample for target binding at the same time. Furthermore, this one-step immobilization strategy greatly simplifies a fabrication process without any chemical modification as well as maintaining activity of biological recognition elements. This system allows specific immobilization of proteins and sensitive detection of targets, which were verified by surface plasmon resonance analysis and successfully applied to electrochemical cyclic voltammetry and impedance spectroscopy upto 0.14 ng/mL HBsAg.
Keywords: hepatitis B virus; Gold-binding polypeptide; fusion protein; electrochemical analysis hepatitis B virus; Gold-binding polypeptide; fusion protein; electrochemical analysis
This is an open access article distributed under the Creative Commons Attribution License (CC BY) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Heo, N.S.; Zheng, S.; Yang, M.; Lee, S.J.; Lee, S.Y.; Kim, H.-J.; Park, J.Y.; Lee, C.-S.; Park, T.J. Label-Free Electrochemical Diagnosis of Viral Antigens with Genetically Engineered Fusion Protein. Sensors 2012, 12, 10097-10108.

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