An amperometric hydrogen peroxide biosensor based on immobilization of horseradish peroxidase on an electrode modified with magnetic dextran microspheres

AbstractA new kind of magnetic dextran microsphere (MDMS) with uniform shape and narrow diameter distribution has been prepared from magnetic iron nanoparticles and dextran. Horseradish peroxidase (HRP) was successfully immobilized on the surface of an MDMS-modified glassy-carbon electrode (GCE), and the immobilized HRP displayed excellent electrocatalytic activity in the reduction of H2O2 in the presence of the mediator hydroquinone (HQ). The effects of experimental variables such as the concentration of HQ, solution pH, and the working potential were investigated for optimum analytical performance. This biosensor had a fast response to H2O2 of less than 10 s and an excellent linear relationship was obtained in the concentration range 0.20 μmol L−1–0.68 mmol L−1, with a detection limit of 0.078 μmol L−1 (S/N = 3) under the optimum conditions. The response showed Michaelis–Menten behavior at larger H2O2 concentrations, and the apparent Michaelis–Menten constant $$ K^{{app}}_{M} $$ was estimated to be 1.38 mmol L−1. Moreover, the selectivity, stability, and reproducibility of the biosensor were evaluated, with satisfactory results. FigureAmperometric response of the biosensor to successive additions of H2O2 and the plot of amperometric response vs. H2O2 concentration

[1]  Han Nim Choi,et al.  Nafion-stabilized magnetic nanoparticles (Fe3O4) for [Ru(bpy)3]2+(bpy = bipyridine) electrogenerated chemiluminescence sensor. , 2005, Chemical communications.

[2]  Zhibing Zhang,et al.  In vitro degradation behavior of microspheres based on cross-linked dextran. , 2006, Biomacromolecules.

[3]  H. Ju,et al.  Direct electrochemistry of horseradish peroxidase immobilized on a colloid/cysteamine-modified gold electrode. , 2000, Analytical biochemistry.

[4]  Hongyuan Chen,et al.  Synthesis and Characterization of Prussian Blue Modified Magnetite Nanoparticles and Its Application to the Electrocatalytic Reduction of H2O2 , 2005 .

[5]  N. Hu,et al.  Direct electron transfer between hemoglobin and pyrolytic graphite electrodes enhanced by Fe(3)O(4) nanoparticles in their layer-by-layer self-assembly films. , 2006, Biophysical chemistry.

[6]  Min-Hung Liao,et al.  Direct Binding and Characterization of Lipase onto Magnetic Nanoparticles , 2003, Biotechnology progress.

[7]  Guo-Li Shen,et al.  Immobilization of horseradish peroxidase to a nano-Au monolayer modified chitosan-entrapped carbon paste electrode for the detection of hydrogen peroxide. , 2003, Talanta.

[8]  M. Yang,et al.  Determination of trace hydrazine by differential pulse voltammetry using magnetic microspheres. , 2001, Talanta.

[9]  W. Hennink,et al.  Controlled release of a model protein from enzymatically degrading dextran microspheres. , 1999, Journal of controlled release : official journal of the Controlled Release Society.

[10]  Y. Chai,et al.  Amperometric Hydrogen Peroxide Biosensor Based on the Immobilization of Horseradish Peroxidase (HRP) on the Layer‐by‐Layer Assembly Films of Gold Colloidal Nanoparticles and Toluidine Blue , 2006 .

[11]  Gang Wang,et al.  Amperometric hydrogen peroxide biosensor with sol-gel/chitosan network-like film as immobilization matrix. , 2003, Biosensors & bioelectronics.

[12]  W. Kunz,et al.  Horseradish peroxidase activity in a reverse catanionic microemulsion. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[13]  Guo-Li Shen,et al.  An amperometric hydrogen peroxide biosensor based on immobilizing horseradish peroxidase to a nano-Au monolayer supported by sol-gel derived carbon ceramic electrode. , 2004, Bioelectrochemistry.

[14]  Jean-Michel Kauffmann,et al.  Amperometric biosensor based on horseradish peroxidase immobilised magnetic microparticles , 2006 .

[15]  Na Wang,et al.  Amperometric third-generation hydrogen peroxide biosensor based on the immobilization of hemoglobin on multiwall carbon nanotubes and gold colloidal nanoparticles. , 2007, Biosensors & bioelectronics.

[16]  L. Deng,et al.  The preparation and catalytically active characterization of papain immobilized on magnetic composite microspheres , 2004 .

[17]  Shujin Li,et al.  A one-step method to construct a third-generation biosensor based on horseradish peroxidase and gold nanoparticles embedded in silica sol–gel network on gold modified electrode , 2005 .

[18]  Guixia Wang,et al.  Electrochemically and catalytically active layer-by-layer films of myoglobin with zirconia formed by vapor-surface sol-gel deposition , 2007 .

[19]  K. Tao,et al.  Preparation of magnetite–dextran microspheres by ultrasonication , 2005 .

[20]  Dongfang Cao,et al.  Electrochemical biosensors utilising electron transfer in heme proteins immobilised on Fe3O4 nanoparticles. , 2003, The Analyst.

[21]  Jing-Juan Xu,et al.  Direct electrochemistry and electrocatalysis of heme proteins immobilized on gold nanoparticles stabilized by chitosan. , 2005, Analytical biochemistry.

[22]  L. Qian,et al.  Composite film of carbon nanotubes and chitosan for preparation of amperometric hydrogen peroxide biosensor. , 2006, Talanta.

[23]  E. Denkbaş,et al.  Magnetic chitosan microspheres: preparation and characterization , 2002 .

[24]  G. Shi,et al.  A water-soluble cationic oligopyrene derivative : Spectroscopic studies and sensing applications , 2009 .

[25]  K. Fernandes,et al.  Covalent immobilisation of horseradish peroxidase onto poly(ethylene terephthalate)–poly(aniline) composite , 2004 .

[26]  Itamar Willner,et al.  Magnetoswitchable electrochemistry gated by alkyl-chain-functionalized magnetic nanoparticles: control of diffusional and surface-confined electrochemical processes. , 2005, Journal of the American Chemical Society.

[27]  Bin Fang,et al.  Fabrication of Fe3O4 Nanoparticles Modified Electrode and Its Application for Voltammetric Sensing of Dopamine , 2005 .

[28]  F. Jiang,et al.  Preparation and properties of soft magnetic particles based on Fe3O4 and hollow polystyrene microsphere composite , 2006 .

[29]  Y. Chai,et al.  A novel and simple biomolecules immobilization method: electro-deposition ZrO2 doped with HRP for fabrication of hydrogen peroxide biosensor. , 2007, Journal of biotechnology.

[30]  Y. Sun,et al.  A Novel Magnetic Affinity Support for Protein Adsorption and Purification , 2001, Biotechnology progress.

[31]  Naifei Hu,et al.  Electrocatalytic Properties of Heme Proteins in Layer‐by‐Layer Films Assembled with SiO2 Nanoparticles , 2004 .

[32]  A. Lewenstam,et al.  Conducting polymer films as model biological membranes: Electrochemical and ion-exchange properties of poly(pyrrole) films doped with asparagine and glutamine , 2006 .

[33]  G. Shen,et al.  Amperometric biosensor with HRP immobilized on a sandwiched nano-Au / polymerized m-phenylenediamine film and ferrocene mediator , 2003, Analytical and bioanalytical chemistry.

[34]  F. Vianello,et al.  A coulometric biosensor to determine hydrogen peroxide using a monomolecular layer of horseradish peroxidase immobilized on a glass surface. , 2007, Biosensors & bioelectronics.

[35]  Li Wang,et al.  A novel hydrogen peroxide sensor based on horseradish peroxidase immobilized on colloidal Au modified ITO electrode , 2004 .

[36]  Huangxian Ju,et al.  Hydrogen peroxide sensor based on horseradish peroxidase-labeled Au colloids immobilized on gold electrode surface by cysteamine monolayer , 1999 .

[37]  K. Hidajat,et al.  Thermosensitive polymer coated nanomagnetic particles for separation of bio-molecules , 2007 .

[38]  Zhuang Li,et al.  Hydrogen peroxide sensor based on horseradish peroxidase immobilized on a silver nanoparticles/cysteamine/gold electrode , 2005, Analytical and bioanalytical chemistry.

[39]  I. Willner,et al.  Switching of directions of bioelectrocatalytic currents and photocurrents at electrode surfaces by using hydrophobic magnetic nanoparticles. , 2005, Angewandte Chemie.