SPECIFIC BINDING ASSAY FOR BIOTIN BASED ON ENZYME CHANNELING WITH DIRECT ELECTRON-TRANSFER ELECTROCHEMICAL DETECTION USING HORSERADISH-PEROXIDASE

被引：25

作者：

WRIGHT, JD ^{[1
]}

RAWSON, KM ^{[1
]}

HO, WO ^{[1
]}

ATHEY, D ^{[1
]}

MCNEIL, CJ ^{[1
]}

机构：

[1] UNIV NEWCASTLE UPON TYNE,SCH MED,DEPT CLIN BIOCHEM,NEWCASTLE TYNE NE2 4HH,TYNE & WEAR,ENGLAND

来源：

BIOSENSORS & BIOELECTRONICS | 1995年 / 10卷 / 05期

关键词：

BIOTIN; CARBON ELECTRODE; DIRECT ELECTRON TRANSFER; GLUCOSE OXIDASE; PEROXIDASE; SPECIFIC BINDING ASSAY;

D O I：

10.1016/0956-5663(95)96895-6

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

A model 'homogeneous' format enzyme channelling specific binding assay for biotin based on peroxide-sensitive horseradish peroxidase mediatorless enzyme electrodes is described. The procedure involved the immobilisation of avidin onto the surface of printed carbon horseradish peroxidase (HRP) enzyme electrodes and the competitive binding of biotin and biotinylated glucose oxidase. Upon addition of glucose, hydrogen peroxide was generated via the glucose oxidase label. Direct electron transfer between the electrodes and HRP resulted in the detection of H2O2 by electroenzymic reduction at +50 mV vs Ag/AgCl. The cathodic current response could be measured in the presence of excess biotinylated glucose oxidase by incorporation of catalase in homogeneous solution to scavenge H2O2 generated in the bulk before it diffused to the electrode surface. The assay showed greatest sensitivity over the range of biotin concentrations 0.07 to 2 mu g ml(-1) in the presence of 10 mu g ml(-1) excess biotinylated glucose oxidase in the bulk solution.

引用

页码：495 / 500

页数：6

共 12 条

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