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Sensors 2009, 9(3), 1794-1806; doi:10.3390/s90301794

Fabricating an Amperometric Cholesterol Biosensor by a Covalent Linkage between Poly(3-thiopheneacetic acid) and Cholesterol Oxidase

1
Department of Chemical Engineering, National Taiwan University/No. 1, Sec. 4, Roosevelt Rd., Taipei 10617, Taiwan
2
Institute of Polymer Science and Engineering, National Taiwan University/No. 1, Sec. 4, Roosevelt Rd., Taipei 10617, Taiwan
*
Author to whom correspondence should be addressed.
Received: 16 December 2008 / Revised: 2 March 2009 / Accepted: 13 March 2009 / Published: 13 March 2009
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Abstract

In this study, use of the covalent enzyme immobilization method was proposed to attach cholesterol oxidase (ChO) on a conducting polymer, poly(3-thiopheneacetic acid), [poly(3-TPAA)]. Three red-orange poly(3-TPAA) films, named electrodes A, B and C, were electropolymerized on a platinum electrode by applying a constant current of 1.5 mA, for 5, 20 and 100 s, respectively. Further, 1-ethyl-3-(3-dimethylamiopropyl)carbodiimide hydrochloride (EDC‧HCl) and N-hydroxysuccinimide (NHS) were used to activate the free carboxylic groups of the conducting polymer. Afterwards, the amino groups of the cholesterol oxidase were linked on the activated groups to form peptide bonds. The best sensitivity obtained for electrode B is 4.49 mA M-1 cm-2,with a linear concentration ranging from 0 to 8 mM, which is suitable for the analysis of cholesterol in humans. The response time (t95) is between 70 and 90 s and the limit of detection is 0.42 mM, based on the signal to noise ratio equal to 3. The interference of species such as ascorbic acid and uric acid increased to 5.2 and 10.3% of the original current response, respectively, based on the current response of cholesterol (100%). With respect to the long-term stability, the sensing response retains 88% of the original current after 13 days. View Full-Text
Keywords: Cholesterol; cholesterol oxidase; covalent; ferrocene; poly(3-thiopheneacetic acid) Cholesterol; cholesterol oxidase; covalent; ferrocene; poly(3-thiopheneacetic acid)
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MDPI and ACS Style

Nien, P.-C.; Chen, P.-Y.; Ho, K.-C. Fabricating an Amperometric Cholesterol Biosensor by a Covalent Linkage between Poly(3-thiopheneacetic acid) and Cholesterol Oxidase. Sensors 2009, 9, 1794-1806.

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