Abstract
In this work, an enzyme biosensor based on the immobilization of horseradish peroxidase (HRP) on SiO2/BSA/Au/thionine/nafion-modified gold electrode was fabricated successfully. Firstly, nafion was dropped on the surface of the gold electrode to form a nafion film followed by chemisorption of thionine (Thi) as an electron mediator via the ion-exchange interaction between the Thi and nafion. Subsequently, the SiO2/BSA/Au composite nanoparticles were assembled onto Thi film through the covalent bounding with the amino groups of Thi. Finally, HRP was immobilized on the SiO2/BSA/Au composite nanoparticles due to the covalent conjugation to construct an enzyme biosensor. The surface topographies of the SiO2/BSA/Au composite nanoparticles were investigated by using scanning electronic microscopy. The stepwise self-assemble procedure of the biosensor was further characterized by means of cyclic voltammetry and chronoamperometry. The enzyme biosensor showed high sensitivity, good stability and selectivity, a wide linear response to hydrogen peroxide (H2O2) in the range of 8.0 × 10-6 ∼ 3.72 × 10-3 mol/L, with a detection limit of 2.0 × 10-6 mol/L. The Michaelies-Menten constant \( K_M^{app} \) value was estimated to be 2.3 mM.
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Abbreviations
- v :
-
Scan rate(mV s-1)
- E :
-
Potential (V)
- I :
-
Current (µA)
- C :
-
Concentration(mM)
- T :
-
Time (s)
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Acknowledgements
This work was supported by the NNSF of China (20675064), the Ministry of Education of China (Project 708073), the Natural Science Foundation of Chongqing City (CSTC-2009BA1003) and High Technology Project Foundation of Southwest University (XSGX02), China.
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Yuan, S., Yuan, R., Chai, Y. et al. Enzyme biosensor based on the immobilization of HRP on SiO2/BSA/Au composite nanoparticles. Appl Biochem Biotechnol 162, 2189–2196 (2010). https://doi.org/10.1007/s12010-010-8993-1
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DOI: https://doi.org/10.1007/s12010-010-8993-1