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Method for preparing biosensor using porous membrane immobilized enzyme

A biosensor, porous membrane technology, applied in biochemical equipment and methods, specific-purpose bioreactors/fermenters, enzymology/microbiology devices, etc. and other problems, to achieve the effect of good stability, simple production method and good reproducibility

Inactive Publication Date: 2009-05-13
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of physical adsorption is that the adsorption amount is small and the stability is poor; the disadvantage of covalent bonding is that the enzyme is easily inactivated during the preparation process.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1: Preparation of glucose oxidase sensor

[0016] First prepare the acrylonitrile-acrylic acid copolymer into a 3% N,N-dimethylformamide solution, then apply 10 μl of the copolymer solution onto the surface of a platinum sheet electrode (3×3 mm), and wait for the solvent to reach 65 After ~85% is evaporated, the electrode is immersed in methanol for 1-5 hours, and then immersed in double-distilled water for 0.5-2 hours to prepare acrylonitrile-acrylic acid copolymer porous membrane. The electrode modified with the polymer porous membrane was immersed in the phosphate buffer (pH6.5, total concentration: 0.1M) containing 8 mg / ml glucose oxidase, with the electrode as the working electrode, the platinum sheet as the auxiliary electrode, saturated glycerin The mercury electrode is used as a reference electrode, and the working electrode is oxidized at a constant potential of 0.60V for 20 minutes to form a glucose sensor based on an acrylonitrile-acrylic acid copolym...

Embodiment 2

[0017] Example 2: Preparation of polyphenol oxidase sensor

[0018] First prepare the acrylonitrile-acrylic acid copolymer into a 2% N,N-dimethylformamide solution, then apply 10 μl of the copolymer solution onto the surface of a glassy carbon electrode (3 mm in diameter), and wait until the solvent is 65- After 85% is evaporated, first immerse the electrode in methanol for 1-5 hours, and then immerse the electrode in twice-distilled water for 0.5-2 hours to prepare the acrylonitrile-acrylic acid copolymer porous membrane. Immerse the electrode modified with the polymer porous membrane into the phosphate buffer (pH6.5, total concentration: 0.1M) containing 2 mg / ml polyphenol oxidase, use the electrode as the working electrode, the platinum wire as the auxiliary electrode, and saturate The calomel electrode was used as a reference electrode, and the working electrode was oxidized at a constant potential of 0.60V for 20 minutes to form a phenol sensor based on acrylonitrile-acry...

Embodiment 3

[0019] Example 3: Preparation of xanthine oxidase sensor

[0020] First prepare the acrylonitrile-acrylic acid copolymer into a 5% N,N-dimethylformamide solution, then apply 10 μl of the copolymer solution onto the surface of a platinum sheet electrode (3×3 mm), and wait for the solvent 65 After ~85% is evaporated, the electrode is immersed in methanol for 1-5 hours, and then immersed in double-distilled water for 0.5-2 hours to prepare acrylonitrile-acrylic acid copolymer porous membrane. Immerse the electrode modified with the polymer porous membrane into the phosphate buffer (pH7.0, total concentration: 0.1M) containing 1 mg / ml xanthine oxidase, use the electrode as the working electrode, the platinum sheet as the auxiliary electrode, and saturate The calomel electrode was used as a reference electrode, and the working electrode was oxidized at a constant potential of 0.60V for 20 minutes to form a xanthine oxidase sensor based on acrylonitrile-acrylic acid copolymer porous...

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Abstract

The invention discloses the method of using the fixing enzyme with the lacunaris film to produce the biosensor and belongs to the method of producing the biosensor with the macromolecule lacunaris film fixing the enzyme. The acrylonitrile-crylic acid assembling content is confected to the N, N-dimethylformamide solution, it is spread on the surface of the floor pole, when the 65-85 percentage of the solution is vaporized, the acrylonitrile-crylic acid assembling lacunaris film is produced using the phase transferring method; then the pole with the decorated polymer is dipped in the enzyme solution, the enzyme is fixed on the acrylonitrile-crylic acid assembling lacunaris film with electrochemistry absorbing method to gain the said biosensor. The produced biosensor has many merits such as the good stability, the high delicacy, the fine recurring ability and needing the little enzyme. The possibility of put in the marketable is great because of the easy producing process. The different functional biosensor can be produced using the same fixing material, so it is fit for the checking of the enzyme bottom material and can be applied in the domain of the medicine, foodstuff and circumstance. It has the good economic benefit.

Description

technical field [0001] The invention belongs to a method for preparing a biosensor by immobilizing enzymes with a polymer porous membrane as a carrier. Background technique [0002] Due to its wide application in medical diagnosis, environmental monitoring, food analysis, etc., biosensor has become one of the frontier research fields. The basic principle is based on the fact that bioactive materials have excellent molecular recognition functions, and have high selectivity and sensitivity to the determination of substances; bioactive substances constitute the molecular recognition part, and the identified information is output as a detectable signal through the information conversion part. . The core of the biosensor is mainly composed of two parts: molecular recognition and information conversion. The molecular recognition part refers to immobilized enzymes, microorganisms, antibodies, cells or tissues, etc., which is the core of the biosensor. Therefore, the immobilizatio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/00C12M1/34
Inventor 薛怀国贺园园王善霞
Owner YANGZHOU UNIV
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