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

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
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Effect test

Example Embodiment

[0039] Example 1

[0040] A method for preparing a biosensor based on a photosensitive biopolymer loaded enzyme includes the following specific steps:

[0041] (1) Preparation of photosensitive biopolymer

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

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

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

Example Embodiment

[0047] Example 2

[0048] A method for preparing a biosensor based on a photosensitive biopolymer loaded enzyme includes the following specific steps:

[0049] (1) Preparation of photosensitive biopolymer

[0050] Combine biological macromolecule dextran (Dex), initiator 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine, small molecule photosensitive monomer cinnamon The acid (CINN) was added dropwise to the dimethyl sulfoxide solvent, and reacted at 60°C for 5 hours to complete the reaction between the biomacromolecule and 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 monomer and initiator; freeze-dry the purified polymer solution to obtain the photosensitive biomacromolecule Dex-CINN;

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

[0052] At room temperature, add 1.0 mg / mL lactate oxidas...

Example Embodiment

[0055] Example 3

[0056] A method for preparing a biosensor based on a photosensitive biopolymer loaded enzyme includes the following specific steps:

[0057] (1) Preparation of photosensitive biopolymer

[0058] The biological macromolecule polyglutamic acid (PGA), initiator 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine, small molecule photosensitive monomer Hydroxyethyl methacrylate (HEMA) was successively added dropwise to the dimethyl sulfoxide solvent and reacted at 25°C for 24 hours to complete the reaction between the biomacromolecule and the photosensitive monomer; the resulting reactant solution was precipitated with ethanol and then transferred Enter the dialysis bag and dialyze for 14 days to remove unreacted monomers and initiators; freeze-dry the purified polymer solution to obtain the photosensitive biological macromolecule PGA-HEMA;

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

[0060] At room temper...

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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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