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Capillary electrochromatography enzyme microreactor based on functionalized graphene and preparation method thereof

A technology of capillary electrochromatography and carboxylated graphene, applied in chemical instruments and methods, enzyme production/bioreactors, biochemical equipment and methods, etc., can solve the problems of complex preparation process, poor separation effect and high production cost , to achieve the effect of simple operation steps, good fixing effect and cost saving

Inactive Publication Date: 2017-12-29
MINZU UNIVERSITY OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This technology is relatively stable, but the preparation process is complicated, the production cost is high, and the separation effect is not good

Method used

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  • Capillary electrochromatography enzyme microreactor based on functionalized graphene and preparation method thereof
  • Capillary electrochromatography enzyme microreactor based on functionalized graphene and preparation method thereof
  • Capillary electrochromatography enzyme microreactor based on functionalized graphene and preparation method thereof

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Embodiment 1

[0059] Capillary electrochromatographic enzyme microreactor preparation method based on functionalized graphene, such as Figure 1-Figure 3 As shown, the specific steps are:

[0060] (1) Pretreatment of the capillary: the inner wall of the capillary is rinsed with 1mol / L sodium hydroxide, deionized water, 0.1mol / L hydrochloric acid, and deionized water for 30 minutes respectively;

[0061] (2) Preparation of capillary electrochromatography of graphene matrix: after blowing dry with nitrogen, feed 5% ethanol solution of 3-aminopropyl-tri(methoxy)-silane to rinse for 10min, so that the inner surface of capillary is bonded with silane Chemical reagent; wash with ethanol for 10min, blow dry with nitrogen; take 2mL of graphene oxide solution with a concentration of 0.5mg / mL, ultrasonicate for 30min (power 100w), add 4mg of carbodiimide (EDC) and 2mg of N-hydroxysuccinate Imide (NHS) activates the carboxyl groups on the graphene oxide, and after vortex shaking for 10 minutes, it is...

Embodiment 2

[0066] Capillary electrochromatographic enzyme microreactor preparation method based on functionalized graphene, such as Figure 1-Figure 3 As shown, the specific steps are:

[0067] (1) Pretreatment of the capillary: the inner wall of the capillary is rinsed with 1mol / L sodium hydroxide, deionized water, 0.1mol / L hydrochloric acid, and deionized water for 30 minutes respectively;

[0068] (2) Preparation of capillary electrochromatography of graphene matrix: after blowing dry with nitrogen, feed 1% ethanol solution of 3-aminopropyl-tri(ethoxy)-silane to rinse for 30min, so that the inner surface of capillary is bonded with silane Chemical reagent; wash with ethanol for 10min, blow dry with nitrogen; take 5mL of graphene oxide solution with a concentration of 5mg / mL, ultrasonicate for 60min (power 150w), add 6mg of carbodiimide (EDC) and 3mg of N-hydroxysuccinyl Imine (NHS) activates the carboxyl groups on the graphene oxide, vortexed for 10 minutes, and rinsed in the capilla...

Embodiment 3

[0073] Capillary electrochromatographic enzyme microreactor preparation method based on functionalized graphene, such as Figure 1-Figure 3 Shown:

[0074] The specific steps are:

[0075] (1) Pretreatment of the capillary: the inner wall of the capillary is rinsed with 1mol / L sodium hydroxide, deionized water, 0.1mol / L hydrochloric acid, and deionized water for 30 minutes respectively;

[0076](2) Preparation of capillary electrochromatography of graphene matrix: after blowing dry with nitrogen, feed 3% ethanol solution of 3-aminopropyl-tri(ethoxy)-silane to rinse for 20min, so that the inner surface of capillary is bonded with silane Chemical reagent; rinse with ethanol for 10min, dry with nitrogen; take 5mL of carboxylated graphene solution with a concentration of 2mg / mL, ultrasonic 20min (power 200w), add 5mg of EDC and 2.5mg of NHS to activate the carboxyl group on the carboxylated graphene , after vortex shaking for 10 minutes, pass into the capillary and rinse for 30 ...

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Abstract

The invention discloses a capillary electrochromatography enzyme microreactor based on functionalized graphene and a preparation method thereof. The preparation method comprises the following steps: (1) pretreating a capillary tube, namely pretreating a capillary tube with alkalis and acids; (2) preparing a graphene substrate based capillary electrochromatography, namely fixing graphene oxide or carboxyl graphene on the inner wall of the capillary tube so as to obtain the graphene substrate based capillary electrochromatography, or preferably fixing the graphene oxide or carboxyl graphene on the inner wall of a whole capillary tube; and (3) preparing an enzyme microreactor, namely introducing a protease solution into one end of the graphene substrate based capillary electrochromatography, performing incubation, and washing with a buffer solution, thereby obtaining the capillary electrochromatography enzyme microreactor based on functionalized graphene. The capillary electrochromatography enzyme microreactor based on functionalized graphene, which is disclosed by the invention, integrates biochemical reactions and separation, and furthermore is high in enzyme immobilization activity, high in separation efficiency for substrates, enzymolysis products and the like, and good in effect.

Description

technical field [0001] The invention belongs to the field of biochemical microreactor production, and in particular relates to a functionalized graphene-based capillary electrochromatographic enzyme microreactor and a preparation method. Background technique [0002] As an important part of the post-gene era, proteomics has become the research frontier and focus in the field of life sciences. In view of the high complexity of protein components and the high disparity in the content of each component, it is particularly critical to develop new protein enzymatic hydrolysis technology and separation and detection technology to realize the rapid separation and efficient identification of various proteins in the system. [0003] As a new type of micro-biochemical reaction tool, the immobilized enzyme microreactor has the advantages of less sample consumption, high enzymolysis efficiency, reusability, and functional assembly of flowers. It is currently used in protein enzymolysis,...

Claims

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Application Information

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IPC IPC(8): B01J19/00C12M1/40C12M3/06
CPCB01J19/0093B01J19/0053B01J2219/0002B01J2219/00792B01J2219/00819B01J2219/0086B01J2219/00916C12M21/18C12M23/16
Inventor 申刚义刘佳张爱芹张蕾
Owner MINZU UNIVERSITY OF CHINA
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