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A kind of dextran-polyhydroxyethyl methacrylate based continuous bed crystal gel separation medium and its preparation method

A technology of poly(hydroxyethyl methacrylate) and separation medium, which is applied in the field of dextran-poly(hydroxyethyl methacrylate)-based continuous bed crystal gel separation medium and its preparation, and can solve the research on dextran-based crystal gel media Lacking, restricting the industrial application of crystal colloidal media, and no research reports, etc., to achieve excellent separation performance, good biocompatibility and safety, and easy scale-up and preparation

Active Publication Date: 2019-04-30
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although scholars at home and abroad have successfully prepared crystal-gel separation media of different matrix materials, and prepared crystal-gel media based on polyhydroxyethyl methacrylate, there is a lack of research on dextran-based crystal-gel media. Dextran derivatives with bonds and hydroxyethyl methacrylate copolymer materials as the matrix crystal gel media have not been studied yet; for dextran-based continuous bed crystal gel media with anion-exchange and hydrophobic dual-mode functional groups, there is also There is no research report, which greatly limits the industrial application of this kind of crystal colloid medium with excellent biosafety performance

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Add 5.0 mL of a polymerizable reaction aqueous solution of glycidyl methacrylate-modified dextran (Dex-MA) and hydroxyethyl methacrylate (HEMA) with a total concentration of 5% (the mass ratio of Dex-MA to HEMA is 1 : 0.1) placed in a column with an inner diameter of 10 mm, added 40 mg ammonium persulfate and 38 mg tetramethylethylenediamine, and carried out crystallization pore formation and copolymerization at a temperature of -12 to 20 °C for 24 h to obtain a continuous bed crystal Gel matrix skeleton; use 15 mL of N,N,N-trimethylvinylbenzyl ammonium chloride solution with a concentration of 0.5 M, and control the temperature at 40-50°C to carry out grafting reaction on the continuous bed crystal gel matrix skeleton. The time is 1 h, and the dextran-polyhydroxyethyl methacrylate copolymer-based continuous bed crystal gel separation medium is obtained. The effective porosity is 88%, the maximum porosity is 92%, and the pore diameter is about 1-200 μm. Protein—The adso...

Embodiment 2

[0023] Put 5.1 mL of Dex-MA and HEMA (mass ratio: 1:0.4) polymerizable reaction aqueous solution with a total concentration of 5% into a column with an inner diameter of 10 mm, add 46 mg of ammonium persulfate and 32 mg of tetramethylethylenediamine, Carry out crystallization pore formation and copolymerization reaction at a temperature of -12-20°C for 24 h to obtain a continuous bed gel matrix skeleton; , control the temperature at 40-50°C, carry out grafting reaction on the matrix skeleton of the continuous bed crystal gel, and the reaction time is 2 h to obtain the dextran-polyhydroxyethyl methacrylate-based copolymer-based continuous bed crystal gel separation medium, which The effective porosity is 88%, the maximum porosity is 96%, the pore diameter is about 1-200 μm, and the adsorption capacity for the model protein - bovine serum albumin is 9 mg / mL.

Embodiment 3

[0025] Put 4.9 mL of 5% Dex-MA and HEMA (mass ratio: 1:1) polymerizable aqueous solution into a column with an inner diameter of 10 mm, add 37 mg of ammonium persulfate and 43 mg of tetramethylethylenediamine, and Carry out crystallization pore formation and copolymerization reaction at a temperature of -12-20°C for 24 h to obtain a continuous bed gel matrix skeleton; use 15 mL of 0.5 M N,N,N-trimethylvinylbenzyl ammonium chloride solution, The temperature is controlled at 40-50°C, and the grafting reaction is carried out on the matrix skeleton of the continuous bed crystal gel. The reaction time is 2 h, and the dextran-polyhydroxyethyl methacrylate-based copolymer-based continuous bed crystal gel separation medium is obtained, which is effective The porosity is 85%, the maximum porosity is 94%, the pore diameter is about 1-200 μm, and the adsorption capacity for the model protein - bovine serum albumin is 6 mg / mL.

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Abstract

The invention discloses a glucan-poly(hydroxyethyl methacrylate) based continuous bed separation medium used in the field of biochemical separation and a preparation method thereof. The pore size range of the separation medium is 1-200 mu m, the porosity is 83-96%, and the adsorption capacity for a model protein such as bovine serum albumin can be up to 5-9 mg / mL cryogel. According to the invention, a matrix scaffold of the provided glucan-poly(hydroxyethyl methacrylate) based continuous bed cryogel separation medium is a copolymerization material of glucan derivatives and hydroxyethyl methacrylate, has favorable biocompatibility and safety and can be reutilized; and functional groups thereof are anion-exchanging and hydrophobic functional dual-mode groups and have excellent separation performance. The preparation method of the glucan-poly(hydroxyethyl methacrylate) based continuous bed cryogel separation medium is simple and easy to implement and easy to realize large-scale preparation, and has wide application prospects in the field of biochemical separation.

Description

technical field [0001] The invention belongs to the technical field of chemical engineering and biological separation, and in particular relates to a dextran-polyhydroxyethyl methacrylate-based continuous bed gel separation medium and a preparation method thereof. Background technique [0002] The continuous bed crystal gel chromatography method is a new type of biological separation technology proposed in recent years. The crystal gel medium has good elasticity in aqueous solution. The crystal gel has super large pores with a pore size ranging from a few microns to hundreds of microns. Microbial cells or fine solid phases are allowed to pass through the bed, and target biomacromolecules can be quickly separated from complex feed fluids containing microbial cells or cell debris at high flow rates. The separation is simple, the mass transfer is rapid, and the separation efficiency is high, so it has important application prospects in the fields of chemical engineering and bio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F285/00C08F251/00C08F226/02C08F220/28B01J20/26B01J20/30
CPCB01J20/264C08F251/00C08F285/00C08F226/02C08F220/281
Inventor 贠军贤耿文超关今韬关怡新姚善泾
Owner ZHEJIANG UNIV OF TECH
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