An HRP based biosensor using sulphonated polyaniline

被引:32
作者
Ngamna, O
Morrin, A
Moulton, SE
Killard, AJ
Smyth, MR
Wallace, GG [1 ]
机构
[1] Univ Wollongong, Intelligent Polymer Res Inst, ARC Ctr Nanostructured Electromat, Wollongong, NSW 2500, Australia
[2] Dublin City Univ, Sch Chem Sci, Dublin 9, Ireland
关键词
polyaniline and derivatives; conducting polymers; HRP biosensors;
D O I
10.1016/j.synthmet.2005.07.259
中图分类号
T [工业技术];
学科分类号
08 ;
摘要
The properties of poly (2-methoxyaniline-5-sulfonic acid) (PMAS), a water-soluble polymer, has been investigated as a potential mediator for biosensors. Addition of poly (L-Lysine) (PLL) to the PMAS renders this material water-insoluble; an important feature for aqueous based biosensors. Characterisation of various PMAS/PLL mixtures showed that a uniform and stable film could reproducibly be fabricated on ITO coated Mylar using 0.1% (w/v) PMAS and 0.05% (w/v) PLL. Immobilisation of enzyme horseradish peroxidase (HRP) onto the PMAS/PLL film was also investigated. Amperometric detection (-0.1 V vs. Ag/AgCl reference electrode) of hydrogen peroxide was investigated using the PMAS/PLL/HRP electrode as the working electrode in a three electrode cell containing phosphate buffer saline (pH 6.4) solution. Amperometric responses were observed upon the addition of hydrogen peroxide. HRP immobilised concentration was optimised to be 0.35% (w/v) at 35 mu L. Loading volume was also studied and optimised to be 35 mu L. The optimal condition for amperometric test was using constant applied potential at -100 mV. Detection limit of the sensor in PBS solution was 0.01 mM H2O2. Calibration curve in PBS pH 6.4 shows a linear response range between 0.01 and 0.1 mM H2O2 with a sensitivity of 24.91 mu A/cm(2) mM(-1) and correlation coefficient of 0.9966.
引用
收藏
页码:185 / 188
页数:4
相关论文
共 10 条
[1]  
BENEDETTO GE, 1996, BIOSENS BIOELECTRON, V11, P1001
[2]   Atrazine analysis using an amperometric immunosensor based on single-chain antibody fragments and regeneration-free multi-calibrant measurement [J].
Grennan, K ;
Strachan, G ;
Porter, AJ ;
Killard, AJ ;
Smyth, MR .
ANALYTICA CHIMICA ACTA, 2003, 500 (1-2) :287-298
[3]   Reactivities of organic phase biosensors. 2. The amperometric behaviour of horseradish peroxidase immobilised on a platinum electrode modified with an electrosynthetic polyaniline film [J].
Iwuoha, EI ;
de Villaverde, DS ;
Garcia, NP ;
Smyth, MR ;
Pingarron, JM .
BIOSENSORS & BIOELECTRONICS, 1997, 12 (08) :749-761
[4]   Amperometric separation-free immunosensor for real-time environmental monitoring [J].
Killard, AJ ;
Micheli, L ;
Grennan, K ;
Franek, M ;
Kolar, V ;
Moscone, D ;
Palchetti, I ;
Smyth, MR .
ANALYTICA CHIMICA ACTA, 2001, 427 (02) :173-180
[5]   Development of an electrochemical flow injection immunoassay (FIIA) for the real-time monitoring of biospecific interactions [J].
Killard, AJ ;
Zhang, SQ ;
Zhao, HJ ;
John, R ;
Iwuoha, EI ;
Smyth, MR .
ANALYTICA CHIMICA ACTA, 1999, 400 :109-119
[6]   Characterisation of horseradish peroxidase immobilisation on an electrochemical biosensor by colorimetric and amperometric techniques [J].
Morrin, A ;
Guzman, A ;
Killard, AJ ;
Pingarron, JM ;
Smyth, MR .
BIOSENSORS & BIOELECTRONICS, 2003, 18 (5-6) :715-720
[7]   Ferrocene-conjugated m-phenylenediamine conducting polymer-incorporated peroxidase biosensors [J].
Mulchandani, A ;
Pan, ST .
ANALYTICAL BIOCHEMISTRY, 1999, 267 (01) :141-147
[8]   Amperometric cholesterol biosensor based on immobilized cholesterol esterase and cholesterol oxidase on conducting polypyrrole films [J].
Singh, S ;
Chaubey, A ;
Malhotra, BD .
ANALYTICA CHIMICA ACTA, 2004, 502 (02) :229-234
[9]   Peroxidase-incorporated sulfonated polyaniline-polycation complexes for electrochemical sensing of H2O2 [J].
Tatsuma, T ;
Ogawa, T ;
Sato, R ;
Oyama, R .
JOURNAL OF ELECTROANALYTICAL CHEMISTRY, 2001, 501 (1-2) :180-185
[10]   Peroxidase-incorporated hydrophilic polythiophene electrode for the determination of hydrogen peroxide in acetonitrile [J].
Tatsuma, T ;
Ariyama, K ;
Oyama, N .
ANALYTICA CHIMICA ACTA, 1996, 318 (03) :297-301