Abstract: 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
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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.
Heo NS, Zheng S, Yang M, Lee SJ, Lee SY, Kim H-J, Park JY, Lee C-S, Park TJ. Label-Free Electrochemical Diagnosis of Viral Antigens with Genetically Engineered Fusion Protein. Sensors. 2012; 12(8):10097-10108.
Heo, Nam Su; Zheng, Shun; Yang, MinHo; Lee, Seok Jae; Lee, Sang Yup; Kim, Hwa-Jung; Park, Jung Youn; Lee, Chang-Soo; Park, Tae Jung. 2012. "Label-Free Electrochemical Diagnosis of Viral Antigens with Genetically Engineered Fusion Protein." Sensors 12, no. 8: 10097-10108.