Abstract
A novel third-generation biosensor for hydrogen peroxide (H2O2) has been constructed based on horseradish peroxidase (HRP) immobilized by the sol–gel (SG) technology on carbon nanotube (CNT)-modified electrode. CNT has good promotion effects on the direct electron transfer between HRP and the electrode surface and the SG network provides a biocompatible microenvironment for enzyme. The immobilized HRP retained its bioelectrocatalytic activity for the reduction of hydrogen peroxide and can respond to the change of concentration of H2O2 rapidly. The heterogeneous electron transfer rate constant was evaluated to be 2.8 ± 0.4 s−1. The amperometric response to H2O2 shows a linear relation in the range from 0.5 to 300 μmol l−1 and a detection limit of 0.1 μmol l−1 (S/N = 3). The K appM value of HRP immobilized on the electrode surface was found to be 1.35 mmol l−1. The biosensor exhibited high sensitivity, rapid response and excellent long-term stability.
Similar content being viewed by others
References
Iijima S (1991) Helical microtubules of graphitic carbon. Nature 354:56–58
Davis JJ, Coleman KS, Azamian BR, Bagshaw CB, Green MLH (2003) Chemical and biochemical sensing with modified single walled carbon nanotubes. Chem Eur J 9:3732–3739
Hamada N, Sawada S, Oshiyama A (1992) New one-dimensional conductors: graphitic microtubules. Phys Rev Lett 68:1579–1581
Davis JJ, Coles RJ, Allen H, Hill O (1997) Protein electrochemistry at carbon nanotube electrodes. J Electroanal Chem 440:279–282
Qi H, Zhang C (2005) Simultaneous determination of hydroquinone and catechol at a glassy carbon electrode modified with multiwall carbon nanotubes. Electroanalysis 17:832–838
Britto PJ, Santhanam KSV, Rubio A, Alonso JA, Ajayan PM (1999) Improved charge transfer at carbon nanotube electrodes. Adv Mater 11:154–157
Wang JX, Li MX, Shi ZJ, Li NQ, Gu ZN (2002) Direct electrochemistry of cytochrome c at a glassy carbon electrode modified with single-wall carbon nanotubes. Anal Chem 74:1993–1997
Zhao GC, Zhang L, Wei XW, Yang ZS (2003) Myoglobin on multi-walled carbon nanotubes modified electrode: direct electrochemistry and electrocatalysis. Electrochem Commun 5:825–829
Wang L, Wang JX, Zhou FM (2004) Direct electrochemistry of catalase at a gold electrode modified with single-wall carbon nanotubes. Electroanalysis 16:627–632
Zhao YD, Bi YH, Zhang WD, Luo QM (2005) The interface behavior of hemoglobin at carbon nanotube and the detection for H2O2. Talanta 65:489–494
Dai YQ, Shiu KK (2004) Glucose biosensor based on multi-walled carbon nanotube modified glassy carbon electrode. Electroanalysis 16:1697–1703
Cai CX, Chen J (2004) Direct electron transfer of glucose oxidase promoted by carbon nanotubes. Anal Biochem 332:75–83
Xu JZ, Zhu JJ, Wu Q, Hu Z, Chen HY (2003) An amperometric biosensor based on the co-immobilization of horseradish peroxidase and methylene blue on a carbon nanotubes modified electrode. Electroanalysis 15:219–224
Yu X, Chattopadhyay D, Galeska I, Papadimitrakopoulos F, Rusling JF (2003) Peroxidase activity of enzymes bound to the ends of single-wall carbon nanotube forest electrodes. Electrochem Commun 5:408–411
Luo XL, Killard AJ, Morrin A, Smyth MR (2006) Enhancement of a conducting polymer-based biosensor using carbon nanotube-doped polyaniline. Anal Chim Acta 575:39–44
Tripathi VS, Kandimalla VB, Ju HX (2006) Amperometric biosensor for hydrogen peroxide based on ferrocene-bovine serum albumin and multiwall carbon nanotube modified ormosil composite. Biosens Bioelectron 21:1529–1535
Zhao YD, Zhang WD, Chen H, Luo QM, Li SFY (2002) Direct electrochemistry of horseradish peroxidase at carbon nanotube powder microelectrode. Sens Actuators B Chem 87:168–172
Cai CX, Chen J (2004) Direct electrochemistry of horseradish peroxidase at a carbon nanotube electrode. Acta Chimi Sin 62:335–340
Qian L, Yang XR (2006) Composite film of carbon nanotubes and chitosan for preparation of amperometric hydrogen peroxide biosensor. Talanta 68:721–727
Ferapontova EE (2004) Direct peroxidase bioelectrocatalysis on a variety of electrode materials. Electroanalysis 16:1101–1111
Gooding JJ (2005) Nanostructuring electrodes with carbon nanotubes: A review on electrochemistry and applications for sensing. Electrochim Acta 50:3049–3060
Wang J (2005) Carbon-nanotube based electrochemical biosensor: a review. Eelectroanalysis 17:7–14
Di JW, Shen CP, Peng SH, Tu YF, Li SJ (2005) A one-step method to construct a therd-generation biosensor based on horseradish peroxidase and gold nanoparticles embedded in silica sol–gel net work on gold modified electrode. Anal Chim Acta 533:196–200
Luo X, Killard AJ, Morrin A, Smyth MR (2006) Enhancement of a conducting polymer-based biosensor usingcarbon nanotube-doped polyaniline. Anal Chim Acta 575:39–44
Song YH, Wang L, Ren CB, Zhu GY, Li Z (2006) A novel hydrogen peroxide sensor based on horseradish peroxidase immobilized in DNA films on a gold electrode. Sens Actuators B Chem 114:1001–1006
Jin W, Brennan JD (2002) Properties and applications of proteins encapsulated within sol–gel derived materials. Anal Chim Acta 461:1–36
Aurobind SV, Amirthalingam KP, Gomathi H (2006) Sol–gel based surface modification of electrodes for electro analysis. Adv Colloid Interface Sci 121:1–7
Wang G, Xu JJ, Chen HY, Lu ZH (2003) Amperometric hydrogen peroxide biosensor with sol–gel/chitosan network-like film as immobilization matrix. Biosens Bioelectron 18:335–343
Rosatto SS, Sotomayor PT, Kubota LT, Gushikem Y (2002) SiO2/Nb2O5 sol–gel as a support for HRP immobilization in biosensor preparation for phenol detection. Electrochim Acta 47:4451–4458
Chen X, Wang BQ, Dong SJ (2001) Amperometric biosensor for hydrogen peroxide based on sol–gel/hydrogel composite thin film. Electroanalysis 13:1149–1152
Lei CX, Hu SQ, Gao N, Shen GL, Yu RQ (2004) An amperometric hydrogen peroxide biosensor based on immobilizing horseradish peroxidase to a nano-Au monolayer supported by sol–gel derived carbon ceramic electrode. Bioelectrochemistry 65:33–39
Liu GD, Lin YH (2006) Amperometric glucose biosensor based on self-assembling glucose oxidase on carbon nanotubes. Electrochem Commun 8:251–256
Tu Y, Di J, Chen X (2005) Study of the nano-size silica sol–gel film as the matrix of chemically modified electrode. J Sol–Gel Sci Technol 33:187–191
Di JW, Bi SP, Zhang M (2004) Third-generation superoxide anion sensor based on superoxide dismutase directly immobilized by sol–gel thin film on gold electrode. Biosens Bioelectron 19:1479–1486
Sun DM, Cai CX, Li XG, Xing W, Lu TH (2004) Direct electrochemistry and bioelectrocatalysis of horseradish peroxidase immobilized on active carbon. J Electroanal Chem 566:415–421
Laviron E (1979) General expression of the linear potential sweep voltammogram in the case of diffusionless electrochemical systems. J Electroanal Chem 101:19–28
Chen XH, Ruan CM, Kong JJ, Deng JQ (2000) Characterization of the direct electron transfer and bioelectrocatalysis of horseradish peroxidase in DNA film at pyrolytic graphite electrode. Anal Chim Acta 412:89–98
Wang BQ, Li B, Deng Q, Dong SJ (1998) Amperometric glucose biosensor based on sol–gel organic-inorganic hybrid material. Anal Chem 70:3170–3174
Kamin RA, Willson GS (1980) Rotation ring-disk enzyme electrode for biocatalysis kinetic studies and characterization of the immobilized enzyme layer. Anal Chem 52:1198–1205
Wang ZH, Liu J, Li QW, Luo GA (2001) The HRP based sol–gel film electrode and the amperometric determination of H2O2. Chin J Anal Chem 29:1482
Acknowledgments
This work was supported by the National Natural Science Foundation of China (no. 20275025).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wang, J., Gu, M., Di, J. et al. A carbon nanotube/silica sol–gel architecture for immobilization of horseradish peroxidase for electrochemical biosensor. Bioprocess Biosyst Eng 30, 289–296 (2007). https://doi.org/10.1007/s00449-007-0126-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00449-007-0126-z