Preparation method of collagen-based glucose fluorescent sensitive membrane

A collagen and glucose technology, applied in the field of fluorescent biosensors and preparation, can solve the problems of unpredictable hyperglycemia and hypoglycemia, reducing patient monitoring initiative, unfavorable long-term monitoring of blood sugar, etc., and achieves fewer preparation processes and safe and reliable preparation process. , the effect of low biological toxicity

Pending Publication Date: 2021-08-13
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, diabetics need to check the blood sugar concentration 5-8 times a day, and frequent pricking of fingers will not only cause tissue damage, but also pose a risk of infection
The daily cumbersome testing times and physical pain make patients resist and greatly reduce th

Method used

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  • Preparation method of collagen-based glucose fluorescent sensitive membrane
  • Preparation method of collagen-based glucose fluorescent sensitive membrane
  • Preparation method of collagen-based glucose fluorescent sensitive membrane

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Prepare mixed enzyme solutions of glucose oxidase and catalase with enzyme activities of 800U / mL and 1600U / mL respectively, and store in a refrigerator at 4°C;

[0029] Cut the collagen base into small pieces of 10mm*10mm and immerse it in the rhodamine 6G solution with a concentration of 0.05mg / mL, take it out after soaking for 80 minutes, and rinse the remaining rhodamine 6G solution on the surface of the fluorescent film with deionized water. Dry at room temperature for later use;

[0030] Immerse the fluorescent film in a glutaraldehyde solution with a mass percentage of 1%, and take it out after soaking for 12 hours;

[0031] Place the activated fluorescent membrane on the surface of a clean glass slide, use a pipette to draw 150 μL of mixed enzyme solution, drop it on the surface of the fluorescent membrane, and add 150 μL of mixed enzyme solution dropwise after 5 hours of crosslinking, and the entire crosslinking The process is repeated 10 times.

Embodiment 2

[0033] Prepare mixed enzyme solutions of glucose oxidase and catalase with enzyme activities of 1000U / mL and 2000U / mL respectively, and store in a refrigerator at 4°C;

[0034] Cut the collagen base into small pieces of 12mm*12mm and immerse it in the rhodamine 6G solution with a concentration of 0.08mg / mL, take it out after soaking for 70 minutes, and rinse the remaining rhodamine 6G solution on the surface of the fluorescent film with deionized water. Dry at room temperature for later use;

[0035] Immerse the fluorescent film in a 0.5% glutaraldehyde solution by mass, and take it out after soaking for 21 hours;

[0036] Place the activated fluorescent membrane on the surface of a clean glass slide, use a pipette to draw 120 μL of mixed enzyme solution, drop it on the surface of the fluorescent membrane, add 120 μL of mixed enzyme solution dropwise after 6 hours of crosslinking, and the entire crosslinking The process was repeated 8 times.

Embodiment 3

[0038] Prepare mixed enzyme solutions of glucose oxidase and catalase with enzyme activities of 1200U / mL and 2400U / mL respectively, and store in a refrigerator at 4°C;

[0039] Cut the collagen base into small pieces of 13mm*13mm and immerse it in the rhodamine 6G solution with a concentration of 0.09mg / mL, take it out after soaking for 60 minutes, and rinse the remaining rhodamine 6G solution on the surface of the fluorescent film with deionized water. Dry at room temperature for later use;

[0040] Immerse the fluorescent film in a glutaraldehyde solution with a mass percentage of 0.2%, and take it out after soaking for 24 hours;

[0041] Place the activated fluorescent membrane on the surface of a clean glass slide, use a pipette to draw 100 μL of mixed enzyme solution, drop it on the surface of the fluorescent membrane, add 100 μL of mixed enzyme solution dropwise after 8 hours of crosslinking, and the entire crosslinking The process was repeated 6 times.

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Abstract

The invention relates to a preparation method of a collagen-based glucose fluorescent sensitive membrane, and belongs to the technical field of fluorescent biosensors and preparation. Rhodamine 6G dye molecules, glucose oxidase and catalase take a collagen substrate material as a matrix, and soluble free molecules can be immobilized onto an insoluble biological substrate, which facilitates repeated use. According to the preparation method, an environment-friendly material with high biocompatibility and low biotoxicity is selected, is a novel fluorescent sensitive membrane, is used for glucose detection, and has the advantages of high enzyme activity, high repeatability, high stability, high selectivity, cleanness, environmental protection and the like. According to the preparation method, technological equipment is simple, preparation processes are few, preparation cost is low, and the fluorescent sensitive membrane is expected to be applied to a wearable continuous monitoring sensor for an organism.

Description

technical field [0001] The invention belongs to the technical field of fluorescent biosensors and preparation, in particular to a method for preparing a collagen-based glucose fluorescent sensitive membrane. Background technique [0002] Non-communicable chronic diseases such as diabetes, hypertension, and hyperlipidemia are the main health threats faced by human beings, especially diabetes caused by defective insulin secretion and impaired insulin action, which is second only to cancer in terms of harm to the human body. The clinical symptoms of diabetic patients are persistent high blood sugar, which will cause chronic damage to the eyes, kidneys, heart, blood vessels and nervous system, leading to cardiovascular and cerebrovascular diseases, renal failure, neuropathy, ketoacidosis and other complications , the patient will face the risk of blindness, disability and amputation, and death. Since the pathogenesis of diabetes is still unclear, the function of pancreatic isle...

Claims

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Application Information

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IPC IPC(8): C12Q1/00G01N33/66G01N21/64
CPCC12Q1/003G01N33/66G01N21/6428G01N2333/904G01N2333/908
Inventor 冯国英彭志清
Owner SICHUAN UNIV
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