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Applied Biochemistry and Biotechnology

, Volume 162, Issue 8, pp 2189–2196 | Cite as

Enzyme biosensor based on the immobilization of HRP on SiO2/BSA/Au composite nanoparticles

  • Shirong Yuan
  • Ruo YuanEmail author
  • Yaqin Chai
  • Ying Zhuo
  • Xia Yang
  • Yali Yuan
Article

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.

Keywords

SiO2/BSA/Au composite nanoparticles Nafion (Nf) Thionine (Thi) HRP Hydrogen peroxide (H2O2

List of symbols

v

Scan rate(mV s-1)

E

Potential (V)

I

Current (µA)

C

Concentration(mM)

T

Time (s)

Notes

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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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Shirong Yuan
    • 1
  • Ruo Yuan
    • 1
    Email author
  • Yaqin Chai
    • 1
  • Ying Zhuo
    • 1
  • Xia Yang
    • 1
  • Yali Yuan
    • 1
  1. 1.Chongqing Key Laboratory of Analytical Chemistry, College of Chemistry and Chemical EngineeringSouthwest UniversityChongqingPeople’s Republic of China

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