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Preparation method of photofixation horse radish peroxidase electric pole and uses thereof

A horseradish peroxidase and electrode technology, which is applied in the direction of material analysis, measuring devices, instruments, etc. by electromagnetic means, can solve the problem of poor stability of horseradish peroxidase electrodes, poor stability of enzyme electrodes, and unfavorable repeated use and other issues, to achieve good stability and reproducibility, good bio-affinity, and long service life

Inactive Publication Date: 2008-03-12
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The problem is that the prepared horseradish peroxidase electrode has poor stability, and the enzyme is prone to shedding during use.
The gels used in the traditional embedding method mainly include polyacrylamide, polyvinyl chloride, methylcellulose, etc. The enzyme electrodes prepared by these materials have poor stability and are easy to fall off from the electrode after long-term use; Poor reversibility, not conducive to repeated use

Method used

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  • Preparation method of photofixation horse radish peroxidase electric pole and uses thereof
  • Preparation method of photofixation horse radish peroxidase electric pole and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Weigh 20g methyl methacrylate, 30g butyl acrylate, 1g vinyl acetate, 0.5g methacrylic acid, fill with N 2 Protected by air, add 1g of benzoyl peroxide, slowly raise the temperature to 95°C, after 2 hours of reaction, check the viscosity, when the viscosity reaches 120mPa·s, cool down, add 0.5g of ethylene glycol dimethacrylate and 5g of phthalate Dioctyl formate and 0.5g benzoin butyl ether are used to make PEA prepolymer. With the above-mentioned PEA prepolymer of 0.1g, be 1: 3 n-butanol: the organic solvent of acetone with 10g mass fraction ratio under dark condition to be formulated into solution, then with the horseradish peroxidase that mass fraction is 5% The equal volumes of the aqueous solutions were mixed and stirred evenly to make a mixed solution. 10 μl of the mixed solution was drip-coated on the surface of the glassy carbon electrode, and irradiated with visible light for 0.1 h at 25°C to polymerize the polymer prepolymer to obtain a photoimmobilized horse...

Embodiment 2

[0029] Weigh 40g methyl methacrylate, 50g butyl acrylate, 3g vinyl acetate, 2g methacrylic acid, fill with N 2 Under gas protection, add 3g of benzoyl peroxide, slowly raise the temperature to 70°C, react for 2 hours, check the viscosity, when the viscosity reaches 120mPa·s, lower the temperature, add 3g of ethylene glycol dimethacrylate and 15g of phthalic acid Dioctyl ester and 5g benzoin butyl ether are used to make PEA prepolymer. With the above-mentioned PEA prepolymer of 0.5g, be 3: 1 n-butanol: the organic solvent of acetone with 10g mass fraction ratio under dark condition to be formulated into solution, then with the horseradish peroxidase that mass fraction is 10% The equal volumes of the aqueous solutions were mixed and stirred evenly to make a mixed solution. 10 μl of the mixed solution was drip-coated on the surface of the graphite electrode, and irradiated with visible light at 20°C for 3 h to polymerize the polymer prepolymer to obtain a photoimmobilized horser...

Embodiment 3

[0031] Weigh 25 g methyl methacrylate, 35 g butyl acrylate, 2 g vinyl acetate, 1 g methacrylic acid, fill with N 2 Under gas protection, add 2g of benzoyl peroxide, slowly raise the temperature to 80°C, react for 2 hours, check the viscosity, when the viscosity reaches 120mPa·s, lower the temperature, add 2g of ethylene glycol dimethacrylate and 10g of phthalic acid Dioctyl ester, 2g benzoin butyl ether to make PEA prepolymer. With the above-mentioned PEA prepolymer of 0.3g, be 1: 1 n-butanol: the organic solvent of acetone with 10g mass fraction ratio under dark condition to be formulated into solution, then with the horseradish peroxidase that mass fraction is 10% The equal volumes of the aqueous solutions were mixed and stirred evenly to make a mixed solution. 10 μl of the mixed solution was drip-coated on the surface of the graphite electrode, and irradiated with visible light at 20 °C for 1 h to polymerize the polymer prepolymer to obtain a photoimmobilized horseradish p...

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Abstract

The present invention provides a photo-fixation horse radish peroxidase preparation method and an application method. 0.5 to 2 portions by weight of polyacrylic ester pre-polymer are dissolved into 100 portions by weight of organic solvent and then mixed with horse radish peroxidase solution to prepare mixed solution. The mixed solution is dipped on an electrode substrate surface, which is illuminated for 0.1h to 3h by visible lights at normal temperature to polymerize the pre-polymer and obtain a photo-fixation horse radish peroxidase electrode. The present invention utilizes characteristics and excellent biological compatibility of polyacrylic esters to prepare the photo-fixation horse radish peroxidase electrode. The photo-fixation horse radish peroxidase electrode prepared by the present invention has the advantages of wide applicable scope and biological catalytic activation for H2O2, organic peroxide and oxygen in aqueous phase, aqueous organic phase and organic phase, quick response, excellent stability and repeatability, long service life, high accuracy, good precision and wide measuring range.

Description

technical field [0001] The invention relates to a photo-immobilized enzyme electrode, in particular to a preparation method and application of a photo-immobilized horseradish peroxidase electrode using polyacrylate as a binder. Background technique [0002] The biosensor developed in the middle and late twentieth century is an interdisciplinary subject interpenetrating and growing from various disciplines such as biology, analytical chemistry, physics, and medicine. It has the advantages of specificity, sensitivity, speed, and accuracy. In the past ten years, a series of valuable biosensors in clinical detection and biochemical analysis have been obtained. The earliest and most widely used biosensor is the enzyme electrode. Enzyme electrodes made by immobilizing enzymes on the surface of electrochemical electrodes, which use enzymes to have special catalytic effects in biochemical reactions, convert the chemical substances consumed or produced during the reaction into elect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
Inventor 高海燕乔毅许芳萍顾伟
Owner JIANGNAN UNIV