Proteolysis micro-fluidic chip based on silica gel oxidized graphene composite membrane and fabrication method of proteolysis micro-fluidic chip

A microfluidic chip and graphene composite technology, which is applied in the direction of enzyme production/bioreactor, can solve the problems of affecting the effect of enzymatic hydrolysis and easy loss, and achieve the effect of simple process, easy production and saving enzymatic hydrolysis time

Inactive Publication Date: 2013-04-03
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The problem is that the enzyme is not fixed by covalent bonds, so it is easy to lose and affect the effect of enzymatic hydrolysis

Method used

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  • Proteolysis micro-fluidic chip based on silica gel oxidized graphene composite membrane and fabrication method of proteolysis micro-fluidic chip
  • Proteolysis micro-fluidic chip based on silica gel oxidized graphene composite membrane and fabrication method of proteolysis micro-fluidic chip
  • Proteolysis micro-fluidic chip based on silica gel oxidized graphene composite membrane and fabrication method of proteolysis micro-fluidic chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1. Proteolysis microfluidic chip based on silica gel graphene oxide composite membrane immobilized with trypsin

[0046] Using computer-aided design software to design the chip structure, the typical design is as follows: figure 1 As shown, it consists of single cross microfluidic channels 2 and 7, solution connection holes 1, 5, 6 and enzymatic solution collection hole 4, and is printed on a transparent film as a mask by using a high-resolution (such as 3600 dpi) laser phototypesetting system. Membrane, the microchannel part is a black line with a width of 40 microns, and the other part is transparent. A layer of positive photoresist ( Shipley S1813 photoresist, Shipley, Marlborough, MA, USA), spin-coating conditions were 3000 rpm for 60 seconds. Then bake at 110 ℃ for 60 seconds to improve the adhesion of the photoresist and remove the residual solvent (baking before exposure), and then cover the mask (including the designed microfluidic structure), using K...

Embodiment 2

[0052] Example 2, proteolysis microfluidic chip based on immobilized protease silica gel graphene oxide composite membrane

[0053] In addition, using the method of the present invention, a proteolysis microfluidic chip based on a silica gel graphene oxide composite membrane immobilized with protease is also prepared. Except for the enzyme immobilization step, others are the same as in Example 1. The specific method is: inject 2 mg / ml 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 1 mg / ml into the modified channel of the silica gel graphene oxide composite membrane. A mixed solution of N-hydroxysuccinimide in 1 ml was used to activate the carboxyl groups of graphene oxide on the channel surface, followed by injection of 50 mmol / L phosphate buffer solution (pH 7.4) containing 2 mg / ml protease at room temperature The reaction was carried out for 4 hours, and the protease was immobilized through covalent bonds to obtain a proteolysis microfluidic chip with a sil...

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Abstract

The invention belongs to the technical field of micro-fluidic chips, and particularly relates to a proteolysis micro-fluidic chip based on a silica gel oxidized graphene composite membrane and a fabrication method of the proteolysis micro-fluidic chip. The fabrication method comprises the steps that chemical oxidization and ultrasonic dispersion are conducted on graphite powder, and an oxidized graphene aqueous solution is obtained; the oxidized graphene aqueous solution is mixed with a silica solution prepared by hydrolyzing n-silicane ethyl ester, and injected into an organic glass micro-fluidic chip channel with the surface subjected to silica gelation treatment; a modification solution is removed after a certain time; a micro-fluidic chip modified with the silica gel oxidized graphene composite membrane is obtained after drying; then, a mixed solution of 1-(3-dimethyl aminopropyl)-3-ethyl carbodiimide hydrochloride and N-hydroxyl succinimide is injected into the channel to allow carboxyl of oxidized graphene on the surface of the channel to be activated; a protease solution such as trypsin is injected to allow protease to be fixed by a covalent bond; and the proteolysis micro-fluidic chip is obtained. A micro-fluidic chip proteolysis reactor has the advantages of short enzymolysis time, low sample consumption, cheap price and the like.

Description

technical field [0001] The invention belongs to the technical field of microfluidic chips, and in particular relates to a proteolysis microfluidic chip based on a silica gel graphene oxide composite film and a preparation method thereof. Background technique [0002] Graphene (graphene) is a new carbonaceous material that is tightly packed into a two-dimensional honeycomb lattice structure by a single layer of carbon atoms. The basic unit of graphite). Since the discovery of single-layer graphene by Geim et al. at the University of Manchester in 2004, [1] , graphene has received extensive attention from scientists all over the world. Graphene has excellent electrical, thermal, and mechanical properties, and is expected to be widely used in high-performance nanoelectronic devices, batteries, electrochemical capacitors, composite materials, field emission materials, gas sensors, and energy storage. [2] . Due to its unique two-dimensional structure and excellent crystallogr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/40
Inventor 陈刚魏邦国包慧敏张鲁雁
Owner FUDAN UNIV
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