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Ionic liquid modified urate oxidase enzyme membrane and preparation method thereof

A urate oxidase enzyme membrane and urate oxidase technology, applied in the field of urate oxidase enzyme membrane and its preparation, can solve the problems of restricting enzyme membrane type electrochemical uric acid sensor, short storage and service life, poor anti-interference ability, etc., to achieve The effect of maintaining and promoting protein activity, improving electrical conductivity, and good electrochemical stability

Inactive Publication Date: 2017-02-08
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The uric acid oxidase enzyme membrane of the enzyme-membrane electrochemical uric acid sensor can be used repeatedly, and the test cost is low; Practical Application of Membrane Electrochemical Uric Acid Sensor

Method used

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  • Ionic liquid modified urate oxidase enzyme membrane and preparation method thereof
  • Ionic liquid modified urate oxidase enzyme membrane and preparation method thereof
  • Ionic liquid modified urate oxidase enzyme membrane and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Immerse the nitrocellulose membrane with an average pore size of 0.45 μm and a thickness of 100 μm in PBS with a pH value of 8.0 and a concentration of 0.1 mol / L for 6 hours, take it out, and use high-purity N 2 Dry it, stick it on the O-shaped rubber ring, and compact it for 3 hours before use; use the phosphate buffer solution PBS with a pH value of 8.0 and a concentration of 0.1mol / L as a solvent to prepare uric acid oxidase activity of 2.5×10 4 The uric acid oxidase solution of U / L, preparation glutaraldehyde volume concentration is the glutaraldehyde solution of 5%, preparation ionic liquid 1-butyl-3-methylimidazole tetrafluoroborate volume concentration is 30% 1- Butyl-3-methylimidazolium tetrafluoroborate solution; respectively take 6 μL of urate oxidase solution, 6 μL of glutaraldehyde solution, and 3 μL of 1-butyl-3-methylimidazolium tetrafluoroborate solution for mixed cross-linking , drop-coated onto the nitrocellulose membrane that has been compacted on the ...

Embodiment 2

[0025] Immerse the nitrocellulose membrane with an average pore size of 0.50 μm and a thickness of 80 μm in a phosphate buffer solution with a pH value of 7.4 and a concentration of 0.1 mol / L for 7 hours, take it out, and use high-purity N 2 Blow dry, stick on the O-shaped rubber ring, compact for 4 hours and set aside; use phosphate buffer solution PBS with a pH value of 7.4 and a concentration of 0.1mol / L as a solvent to prepare uric acid oxidase activity of 3.0×10 4 The uric acid oxidase solution of U / L, preparation glutaraldehyde volume concentration is the glutaraldehyde solution of 3%, preparation ionic liquid 1-butyl-3-methylimidazole tetrafluoroborate volume concentration is 25% 1- Butyl-3-methylimidazolium tetrafluoroborate solution; respectively take 6 μL of urate oxidase solution, 6 μL of glutaraldehyde solution, and 3 μL of 1-butyl-3-methylimidazolium tetrafluoroborate solution for mixed cross-linking , drop-coated onto the nitrocellulose membrane that has been com...

Embodiment 3

[0028] Immerse the nitrocellulose membrane with an average pore size of 0.8 μm and a thickness of 100 μm in a phosphate buffer solution with a pH value of 7.0 and a concentration of 0.1 mol / L for 6 hours, take it out, and use high-purity N 2 Dry it, stick it on the O-shaped rubber ring, and compact it for 3 hours before use; use the phosphate buffer solution PBS with a pH value of 7.0 and a concentration of 0.1mol / L as a solvent to prepare uric acid oxidase activity of 5.0×10 4 The uric acid oxidase solution of U / L, preparation glutaraldehyde volume concentration is the glutaraldehyde solution of 5%, preparation ionic liquid 1-butyl-3-methylimidazole tetrafluoroborate volume concentration is 20% 1- Butyl-3-methylimidazolium tetrafluoroborate solution; respectively take 6 μL of urate oxidase solution, 6 μL of glutaraldehyde solution, and 3 μL of 1-butyl-3-methylimidazolium tetrafluoroborate solution for mixed cross-linking , drop-coated onto the nitrocellulose membrane that has...

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Abstract

The invention provides an ionic liquid modified urate oxidase enzyme membrane and a preparation method thereof, and belongs to the technical field of an electrochemical biosensor and preparation thereof. The enzyme membrane uses a nitrocellulose membrane as a substrate membrane, and is loaded with urate oxidase, ionic liquid of 1-butyl-3-methylimidazolium tetrafluoroborate and crosslinking agents of glutaraldehyde. The preparation method of the enzyme membrane comprises the following steps that the nitrocellulose membrane is soaked into a PBS (phosphate buffer solution) and is then dried through being blown by N2; next, the nitrocellulose membrane is fixed by an O ring; the urate oxidase, glutaraldehyde and 1-butyl-3-methylimidazolium tetrafluoroborate solution is subjected to mixed crosslinking; next, a mixed solution is taken and is dripped and coated onto the nitrocellulose membrane; after the drying, the enzyme membrane provided by the invention is obtained. The ionic liquid modified urate oxidase enzyme membrane has the advantages that the ionic liquid effectively improves the electric conductivity of the enzyme membrane; the current response time is shortened; the enzyme membrane is used for uric acid detection, and has the advantages of wide linear range, high sensitivity and good stability; in addition, the method is simple; the implementation is easy; the cost is low.

Description

technical field [0001] The invention belongs to the technical field of electrochemical biosensors and preparation thereof, in particular to a urate oxidase enzyme film modified with ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate and a preparation method. Background technique [0002] Electrochemical biosensor is a hotspot in the field of sensor research because of its advantages of accurate analysis, rapidity, good selectivity and high sensitivity. Due to the importance of uric acid detection in clinical medicine, research on electrochemical uric acid sensors has attracted much attention. The uric acid oxidase enzyme membrane of the enzyme-membrane electrochemical uric acid sensor can be used repeatedly, and the test cost is low; Practical application of membrane-type electrochemical uric acid sensor. In order to solve the above problems, people mainly carry out research work from the aspects of screening new immobilization matrix materials, adding suitable nan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
CPCG01N27/3272
Inventor 杨文胜曹芮陈旭
Owner BEIJING UNIV OF CHEM TECH
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