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Glucose biosensing membrane, glucose oxidase and preparation method thereof

A glucose oxidase and sediment technology, applied in the field of glucose biosensing membranes, can solve problems affecting product consistency, high viscosity of glucose oxidase solution, etc., and achieve the effect of improving sensitivity and maintaining catalytic oxidation performance

Active Publication Date: 2022-04-19
SUZHOU ZHONGXING MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

What's more serious is that with the increase of use time, there will be more and more chemical cross-linking reactions, and the viscosity of the glucose oxidase solution will also increase, which will seriously affect the consistency of the product.

Method used

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  • Glucose biosensing membrane, glucose oxidase and preparation method thereof
  • Glucose biosensing membrane, glucose oxidase and preparation method thereof
  • Glucose biosensing membrane, glucose oxidase and preparation method thereof

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Effect test

preparation example Construction

[0041] The preparation method of glucose oxidase, comprises the steps:

[0042] A), the deprosylation of glucose oxidase:

[0043] A1), dissolving glucose oxidase in 3~7mol / L ammonium sulfate solution, culturing for 1~4 hours, and then performing centrifugation at a speed of 10000~15000 rpm for 10~30 minutes; preferably, ammonium sulfate The concentration is 4.5~6mol / L;

[0044] A2) Discard the upper layer of centrifuged liquid, dissolve the precipitate in 0.1~2mol / L sodium acetate solution, and slowly add the sodium acetate solution into 3~7mol / L ammonium sulfate solution, the pH of the solution is 1.4, and incubate for 1~4 hours , and then centrifuged again at a speed of 10,000 to 15,000 revolutions per minute for 10 to 30 minutes, repeating this step several times; preferably, the concentration of ammonium sulfate is 4.5 to 6 mol / L;

[0045] A3), discard the supernatant centrifugate, wash the precipitate with phosphate buffer solution of 12~36mmol / L (preferably, 18~27mmol...

Embodiment S1

[0064] (1), the deprosylation of glucose oxidase:

[0065] Dissolve 2 g of glucose oxidase in a 5.6 mol / L ammonium sulfate solution with a pH of 1.4 and incubate for 2 hours; then, pour the obtained ammonium sulfate solution into a centrifuge tube and centrifuge at a speed of 1300 rpm for 20 minutes; Discard the supernatant, and dissolve the precipitate in 1 mol / L sodium acetate; slowly add the sodium acetate solution to a 5.6 mol / L ammonium sulfate solution with a pH of 1.4, and after incubating for 2 hours, pour the obtained solution into a centrifuge tube Centrifuge at a speed of 13,000 rpm for 20 minutes; discard the supernatant, and dissolve the precipitate in 1 mol / L sodium acetate; slowly add the sodium acetate solution to a 5.6 mol / L ammonium sulfate solution with a pH of 1.4, Cultivate for 2 hours, then pour the resulting solution into a centrifuge tube, and centrifuge at a speed of 13,000 rpm for 20 minutes; discard the supernatant and wash with 20 mmol / L phosphate b...

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Abstract

The invention discloses a glucose biosensing membrane, a glucose oxidase and a preparation method thereof. The preparation method of the recombinant glucose oxidase comprises the following steps: A), aposynthesis of the glucose oxidase; B), a metal complex and black Coupling of phosphorus nanosheets and prosthetic groups of glucose oxidase; C), recombination of glucose oxidase. The present invention recombines glucose oxidase onto black phosphorus nanosheets containing electron transfer relay points, and successfully develops a glucose biosensing membrane with extremely low redox potential; the glucose biosensor prepared from the glucose biosensing membrane, It not only maintains the catalytic oxidation performance of glucose, but also realizes the monitoring of glucose at a very low potential (‑100~0 mV), which significantly improves the sensitivity, accuracy, stability and specificity of glucose biosensors sex.

Description

technical field [0001] The invention relates to a glucose biosensing film, a glucose oxidase and a preparation method thereof. Background technique [0002] The continuous glucose monitoring system developed rapidly in recent years is favored by more and more diabetic patients due to its convenient use and real-time monitoring. As the core component of the continuous glucose monitoring system, the performance of the glucose biosensor directly determines the performance and service life of the continuous glucose monitoring system. Glucose biosensors used in existing continuous glucose monitoring systems can be divided into two categories. One is the indirect monitoring of glucose by electrochemically detecting hydrogen peroxide generated during the catalytic oxidation of glucose by glucose oxidase, such as Dekang's Dexcom G5 and G6 and Medtronic's Guardian and iPro 2. They rely on oxygen in interstitial fluid to monitor glucose, and the oxygen content in interstitial fluid ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
CPCG01N27/3272
Inventor 沈薇
Owner SUZHOU ZHONGXING MEDICAL TECH CO LTD
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