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. 2009;9(6):4635-48.
doi: 10.3390/s90604635. Epub 2009 Jun 12.

Immobilization of HRP in Mesoporous Silica and Its Application for the Construction of Polyaniline Modified Hydrogen Peroxide Biosensor

Chien-Chung Chen  1 , Jing-Shan Do, Yesong Gu
Affiliations

Immobilization of HRP in Mesoporous Silica and Its Application for the Construction of Polyaniline Modified Hydrogen Peroxide Biosensor

Chien-Chung Chen et al. Sensors (Basel). 2009.

Abstract

Polyaniline (PANI), an attractive conductive polymer, has been successfully applied in fabricating various types of enzyme-based biosensors. In this study, we have employed mesoporous silica SBA-15 to stably entrap horseradish peroxidase (HRP), and then deposited the loaded SBA-15 on the PANI modified platinum electrode to construct a GA/SBA-15(HRP)/PANI/Pt biosensor. The mesoporous structures and morphologies of SBA-15 with or without HRP were characterized. Enzymatic protein assays were employed to evaluate HRP immobilization efficiency. Our results demonstrated that the constructed biosensor displayed a fine linear correlation between cathodic response and H(2)O(2) concentration in the range of 0.02 to 18.5 mM, with enhanced sensitivity. In particular, the current approach provided the PANI modified biosensor with improved stability for multiple measurements.

Keywords: SBA-15; biosensor; horseradish peroxidase; hydrogen peroxide; polyaniline.

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Figures

Figure 1.
Figure 1.
XRD pattern of (a) SBA-15 and (b) SBA-15(HRP).
Figure 2.
Figure 2.
Nitrogen adsorption and desorption isotherms of SBA-15 before (●) and after (▲) the immobilization of HRP. Inset indicated the pore size distribution of SBA-15 (●) and SBA-15(HRP) (▲).
Figure 3.
Figure 3.
The TEM image of SBA-15.
Figure 4.
Figure 4.
The SEM images of the surfaces of (a) PANI/Pt, (b) SBA-15/PANI/Pt, and (c) GA/SBA-15(HRP)/PANI/Pt electrodes.
Figure 5.
Figure 5.
The cyclic voltammograms of: (a) PANI/Pt, (b) GA/SBA-15/PANI/Pt, and (c) GA/SBA-15(HRP)/PANI/Pt in response to 18.5 mM of H2O2 in 0.1 M phosphate buffer (pH 6.2) with a scan rate of 20 mV/s.
Figure 6.
Figure 6.
The linear calibrations of (a) PANI/Pt, (b) GA/SBA-15/PANI/Pt, and (c) GA/SBA-15(HRP)/PANI/Pt with various concentration of H2O2 in 0.1 M phosphate buffer (pH 6.2). The data were collected from three separate experiments on three individual sensors.
Figure 7.
Figure 7.
The current responses of (a) PANI/Pt, (b) GA/HRP/PANI/Pt, (c) SBA-15(HRP)/PANI/Pt, and (d) GA/SBA-15(HRP)/PANI/Pt electrodes to 1.96 mM H2O2 in a 16-day period with multiple measurements. The data were collected from three separate experiments on the same sensor.
See this image and copyright information in PMC

References

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