Fast protease microreactor and its production

A technology of proteolytic enzyme and reactor, which is applied in the field of miniature fast proteolytic enzyme reactor and its preparation, can solve the problems of high price and limited application of active monomers, and achieve good permeability and bioaffinity, and application The effect of high potential and increased reaction speed

Inactive Publication Date: 2007-06-20
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the above-mentioned monolithic substrates can make efficient enzyme reactors, the price of these active monomers is relatively expensive, which limits their application

Method used

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  • Fast protease microreactor and its production
  • Fast protease microreactor and its production
  • Fast protease microreactor and its production

Examples

Experimental program
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Effect test

Embodiment 1

[0035]1) Preparation of monolithic matrix: as shown in Figure 1, select glycidyl methacrylate (GMA), acrylamide and ethylidene dimethacrylic acid (EDMA) as monomers to synthesize monolithic matrix in capillary. The inner wall of the capillary is pre-modified by a silane reagent (3-trimethoxysilyloxypropyl methacrylate, γ-MAPS) with a double bond functional group, and monomers are polymerized in it and fixed on the capillary wall by chemical bonds. Specifically, such a porous material is polymerized in-situ in a capillary to form an integral matrix enzyme reactor and applied to hydrolysis experiments of various substrates. Weigh 90mg of GMA, 60mg of acrylamide and 150mg of EDMA, dissolve in 700mg of cyclohexanol / dodecanol (mass ratio 85 / 15) mixed solvent, heat to above 50°C, oscillate and mix, cool to room temperature, add content AIBN accounting for 1% of the total mass ratio of the monomer was used as an initiator, and after 10 minutes of nitrogen gas flow, ultrasonic degassi...

Embodiment 2

[0040] 1) Preparation of monolithic matrix: as shown in Figure 1, select glycidyl methacrylate (GMA), acrylamide and ethylidene dimethacrylic acid (EDMA) as monomers to synthesize monolithic matrix in capillary. The inner wall of the capillary is pre-modified by a silane reagent (3-trimethoxysilyloxypropyl methacrylate, γ-MAPS) with a double bond functional group, and monomers are polymerized in it and fixed on the capillary wall by chemical bonds. Specifically, such a porous material is polymerized in-situ in a capillary to form an integral matrix enzyme reactor and applied to hydrolysis experiments of various substrates. Weigh 90mg of GMA, 60mg of acrylamide and 150mg of EDMA, dissolve in a mixed solvent of 700mg of cyclohexanol / dodecanol (mass ratio 50 / 50), heat to 80°C, shake and mix well, after cooling to room temperature, add the content of AIBN with a total monomer mass ratio of 2% was used as an initiator, and after nitrogen gas flow for 10 minutes, ultrasonic degassin...

Embodiment 3

[0045] 1) Preparation of monolithic matrix: as shown in Figure 1, select glycidyl methacrylate (GMA), acrylamide and ethylidene dimethacrylic acid (EDMA) as monomers to synthesize monolithic matrix in capillary. The inner wall of the capillary is pre-modified by a silane reagent (3-trimethoxysilyloxypropyl methacrylate, γ-MAPS) with a double bond functional group, and monomers are polymerized in it and fixed on the capillary wall by chemical bonds. Specifically, such a porous material is polymerized in-situ in a capillary to form an integral matrix enzyme reactor and applied to hydrolysis experiments of various substrates. Weigh 90mg of GMA, 60mg of acrylamide and 150mg of EDMA, dissolve in a mixed solvent of 700mg of n-propanol / 1,4-butanediol (mass ratio 60 / 40), heat to above 50°C, shake and mix well, and cool to room temperature , adding AIBN with a content of 1% of the total mass ratio of monomers as an initiator, and after passing nitrogen gas for 10 minutes, ultrasonic de...

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Abstract

A fast micro-protease reactor is prepared by taking multiple polymer monomers in capillaries, in-situ polymerizing in higher alcohol mixed solution to obtain continuous homogenous porous polymer substrate, activating while modifying for group and binding trypsase molecule on substrate to obtain the final product. It has fast high-activity protease molecule, better stability and less side reactions.

Description

technical field [0001] The invention relates to a miniature fast proteolytic enzyme reactor and its preparation, which is used for rapid enzymolysis of protein by fixing trypsin on a porous monolithic matrix. The continuous and uniform porous structure of the reactor can significantly improve the mass transfer and convection of the substrate in the substrate, and the prepared enzyme reactor has high reactivity, and greatly improves the reaction speed of the enzyme and the substrate. Background technique [0002] With the completion of the human genome, the study of the proteome has gradually become a research hotspot. In order to meet the requirements of sequence analysis and rapid identification of a large number of proteins, it is necessary to develop fast and efficient protein research methods. For the identification of proteins, proteolytic enzymes must be used to hydrolyze proteins. At present, the most used method is to directly add trypsin to the aqueous solution to...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12N11/08
Inventor 张玉奎段继诚张丽华张维冰梁振张洁
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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