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Fixing method of high-activity glucose oxidase

A technology of glucose oxidase and immobilization method, which is applied in the field of immobilization of high-activity glucose oxidase, can solve problems such as complex operation, easy inactivation of enzyme molecules, and harsh reaction conditions, and achieve simple process, good biocompatibility, and improved The effect of catalytic activity

Active Publication Date: 2019-01-25
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But because the reaction conditions of this method are harsh, the operation is complicated, and the enzyme molecule is easily inactivated, so the specific activity of the immobilized enzyme is generally not high, and some may even cause some changes in the properties of the enzyme (such as the base of the enzyme). object specificity)

Method used

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  • Fixing method of high-activity glucose oxidase
  • Fixing method of high-activity glucose oxidase
  • Fixing method of high-activity glucose oxidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Weigh 1.2g of HEMA, 0.4g of NVP, add 0.4% of photoinitiator Darocur 1173, 0.3% of cross-linking agent EGDMA into it, stir evenly to obtain a mixed solution; take 100 μL of the mixed solution, pour it into Add 10 mg of glucose oxidase, and sonicate to dissolve the enzyme completely. Take 7μL of the enzyme-dissolved solution and drop-coat it on the surface of the Pt electrode at a wavelength of 365nm and an intensity of 30mW / cm 2 Cured under ultraviolet light for 50 minutes, after curing, put the Pt electrode into PBS solution at room temperature for hydration, remove unreacted monomers, and obtain glucose oxidase-modified electrodes.

Embodiment 2

[0038] Weigh 1g of HPMA, 0.6g of NVP, add 0.4% photoinitiator Darocur 1173, 0.3% cross-linking agent EGDMA to it, stir well to obtain a mixed solution; take 100 μL of the mixed solution, add 10mg of glucose oxidase, ultrasonic treatment to dissolve the enzyme completely. Take 7μL of the enzyme-dissolved solution and drop-coat it on the surface of the Pt electrode at a wavelength of 365nm and an intensity of 30mW / cm 2 Cured under ultraviolet light for 50 minutes, after curing, put the Pt electrode into PBS solution at room temperature for hydration, remove unreacted monomers, and obtain glucose oxidase-modified electrodes.

[0039] figure 2 The current response to glucose of the sensor prepared by using HEMA and NVP as the photosensitive resin curing enzyme of the hydrophilic monomer in Example 1 of the present invention; image 3 The current response of the sensor made for the photosensitive resin solidifying enzyme of the embodiment 2 of the present invention using HPMA an...

Embodiment 3

[0041] Weigh 0.6g of HPMA, 1g of NVP, 0.4g of VTEO, add 0.4% of photoinitiator Darocur1173, 0.3% of cross-linking agent EGDMA, stir well to obtain a mixed solution; take 100 μL of mixed solution , adding 10 mg of glucose oxidase to it, and ultrasonically dissolving the enzyme. Take 7μL of the enzyme-dissolved solution and drop-coat it on the surface of the Pt electrode at a wavelength of 365nm and an intensity of 30mW / cm 2 Cured under ultraviolet light for 50 minutes, after curing, put the Pt electrode into PBS solution at room temperature for hydration, remove unreacted monomers, and obtain glucose oxidase-modified electrodes.

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Abstract

The invention discloses a fixing method of high-activity glucose oxidase. Glucose oxidase is added into a water soluble photosensitive resin monomer solution containing a silane coupling agent, the glucose oxidase is buried in the net structure of the cured photosensitive resin through photopolymerization. The oxygen permeation rate of the cured resin film is greatly increased by adding the silanecoupling agent into the water soluble photosensitive resin, and accordingly the activity of fixed glucose oxidase is effectively improved. The fixing method steps are simple, all reagents applied inthe method have the good biocompatibility, and the prepared sensor has the good detection effect and high response speed on glucose.

Description

technical field [0001] The invention relates to a method for immobilizing highly active glucose oxidase, which belongs to the technical field of immobilized enzymes. Background technique [0002] Diabetes mellitus is a metabolic disease characterized by hyperglycemia caused by defects in insulin secretion in the body or insulin resistance in tissue cells. The main clinical manifestations of diabetes are "three excesses and one deficiency", that is, polydipsia, polyuria, polyphagia and weight loss. Diabetes can cause various complications. Acute complications include hypoglycemia, ketoacidosis, and nonketotic hyperosmolar coma. Chronic long-term complications include cardiovascular disease, chronic renal failure, retinopathy, neuropathy, microangiopathy, and diabetic foot. The main cause of chronic complications is long-term high blood sugar. Early diagnosis and treatment of diabetic patients, effective control of blood sugar concentration and good living habits can effec...

Claims

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Application Information

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IPC IPC(8): G01N27/327
CPCG01N27/3271
Inventor 王志飞白林贺祖茂
Owner SOUTHEAST UNIV