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Method based on photosensitive biomacromolecule-loaded enzyme to prepare biosensor

A biopolymer and biosensor technology, applied in the field of polymer materials and electrochemical sensors, achieves the effects of simple operation, long analysis time and good biocompatibility

Active Publication Date: 2017-01-11
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the photosensitive biopolymer-enzyme self-assembled nanoparticles prepared by self-assembly and used to construct enzyme biosensors have not been reported yet.

Method used

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  • Method based on photosensitive biomacromolecule-loaded enzyme to prepare biosensor
  • Method based on photosensitive biomacromolecule-loaded enzyme to prepare biosensor
  • Method based on photosensitive biomacromolecule-loaded enzyme to prepare biosensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] A method for preparing a biosensor based on photosensitive biopolymer-loaded enzymes, comprising the following specific steps:

[0041] (1) Preparation of photosensitive biopolymer

[0042] Biomacromolecule chitosan (CS), initiator 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, small molecule photosensitive monomer caffeic acid (CA) were added drop by drop Add 1% acetic acid solution and react at 0°C for 48 hours to completely react biomacromolecules and photosensitive monomers; the resulting reactant solution is precipitated with precipitant ethanol and transferred to a dialysis bag for dialysis for 3 days to remove unreacted monomers and Initiator; freeze-dry the purified polymer solution to obtain photosensitive biomacromolecule CS-CA;

[0043] (2) Preparation of enzyme-loaded self-assembled nanoparticles

[0044] At room temperature, add 0.1 mg / mL glucose oxidase (GOx) solution dropwise into an equal volume of 0.5 mg / mL photosensitive biomacromolecul...

Embodiment 2

[0048] A method for preparing a biosensor based on photosensitive biopolymer-loaded enzymes, comprising the following specific steps:

[0049] (1) Preparation of photosensitive biopolymer

[0050] Biomacromolecule dextran (Dex), initiator 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine, small molecule photosensitive monomer cinnamon The acid (CINN) was added dropwise into the dimethyl sulfoxide solvent, and reacted at 60°C for 5 hours to completely react the biomacromolecule with the photosensitive monomer; the resulting reactant solution was precipitated with precipitant ethanol and transferred to a dialysis bag for dialysis for 7 days To remove unreacted monomers and initiators; freeze-dry the purified polymer solution to obtain the photosensitive biomacromolecule Dex-CINN;

[0051] (2) Preparation of enzyme-loaded self-assembled nanoparticles

[0052] At room temperature, 1.0 mg / mL lactate oxidase (LOx) solution was added dropwise t...

Embodiment 3

[0056] A method for preparing a biosensor based on photosensitive biopolymer-loaded enzymes, comprising the following specific steps:

[0057] (1) Preparation of photosensitive biopolymer

[0058] Biomacromolecule polyglutamic acid (PGA), initiator 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine, small molecule photosensitive monomer Hydroxyethyl methacrylate (HEMA) was added dropwise into the dimethyl sulfoxide solvent, and reacted at 25°C for 24 hours to completely react the biomacromolecule with the photosensitive monomer; Dialyze into a dialysis bag for 14 days to remove unreacted monomers and initiators; freeze-dry the purified polymer solution to obtain photosensitive biomacromolecule PGA-HEMA;

[0059] (2) Preparation of enzyme-loaded self-assembled nanoparticles

[0060] At room temperature, 10 mg / mL horseradish peroxidase (HRP) solution was added dropwise to 10 times its volume of 100 mg / mL photosensitive biomacromolecule PGA-...

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Abstract

The invention discloses a method based on photosensitive biomacromolecule-loaded enzyme to prepare a biosensor. The method includes three steps: preparing photosensitive biomacromolecules; preparing enzyme-loaded self-assembly nanoparticles; preparing an enzyme biosensor. Through self-assembly of the photosensitive biomacromolecules and enzyme, efficient loading of the enzyme can be realized, and activity of the enzyme can be maintained; a self-assembly particle polymerization network acquired through ultraviolet crosslinking can enhance stability of a coating and can effectively prevent the enzyme from leaking from the surface of a sensor matrix, and the enzyme biosensor has the advantages of high specificity, high stability and wide detection range; by combining self-assembly technology, photocuring technology and an electrochemical sensor, the biosensor can be widely applied in the field of food safety, biomedicine and environment protection monitoring.

Description

technical field [0001] The invention relates to the field of macromolecular materials and electrochemical sensors, in particular to a method for realizing effective loading of biocatalyzed enzymes through photosensitive biopolymers and using it for preparing enzyme biosensors. Background technique [0002] As a biocatalyst, enzyme has the characteristics of fast reaction speed, mild reaction conditions and high selectivity. However, in actual use, most enzymes are more sensitive to the environment. Under the influence of external conditions such as high temperature, strong acid, strong alkali, ultraviolet rays, heat, metal salts, etc., the secondary and tertiary structures of the enzyme will change, resulting in the loss of enzyme activity. , thus losing its functionality. [0003] As one of the most widely studied biosensors, enzyme biosensors use enzymes as biological sensitive elements to capture the reaction between the target and the sensitive element through various p...

Claims

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Application Information

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IPC IPC(8): G01N27/30C08F251/00C08F283/04
CPCC08F251/00C08F283/04G01N27/30
Inventor 刘晓亚许升吴倩王艺晨赵伟罗静朱叶魏玮
Owner JIANGNAN UNIV
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