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Glucosan-based cryogel microsphere separating medium and preparation method thereof

A technology for dextran-based crystals and separation media, which is applied in chemical instruments and methods, other chemical processes, etc., can solve the problems of lack of research on crystalline colloidal granular media, limitation of industrial application of crystalline colloidal media, difficulties, etc., and achieve a good biological phase. Capacitance and safety, uniform and controllable particle size, and large pore size

Active Publication Date: 2017-05-10
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is a lack of research on dextran-based crystal gel granular media. The main reason is that it is difficult to shape dextran by traditional methods and graft it into granular crystal gels, which greatly limits the use of this type of crystal gel media. industrial applications

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Use 4 mL of an aqueous solution of polymerizable monomers containing glycidyl methacrylate-modified dextran (Dex-GMA) with a total concentration of 5% as the aqueous phase solution; use ethyl heptanoate as the oil phase solution, and set the flow rate in the water phase Microfluidic focusing was performed at a flow rate of 0.3 cm / s and an oil phase flow rate of 9 cm / s to form polymerizable droplets with a particle size of 520-1900 μm in 40 mL of ethyl heptanoate; the polymerizable droplets were crystallized to Pore ​​and polymerization reaction to obtain the matrix skeleton of dextran-based crystalloid microspheres; use 30 mL of AMPSA solution with a concentration of 0.25 M and control the temperature at 40-55 °C to carry out grafting reaction on the matrix skeleton of dextran-based crystalloid microspheres , the reaction time was 2 h, and the separation medium of dextran-based crystal gel microspheres was obtained. The particle size was 520-1900 μm, the pore size was 1-...

Embodiment 2

[0024] 4 mL of polymerizable monomer aqueous solution containing 5% total concentration of Dex-GMA was used as the water phase solution; n-hexane was used as the oil phase solution, and the flow rate of the water phase was 0.3 cm / s, and the flow rate of the oil phase was 6 cm / s. Microfluidic focusing into droplets, forming polymerizable droplets with a particle size of 500-950 μm in 250 mL of n-hexane; the polymerizable droplets undergo crystallization and polymerization reactions to obtain the matrix skeleton of dextran-based crystalloid microspheres; Using 30 mL of AMPSA solution with a concentration of 1 M, the temperature was controlled at 40-55 °C to carry out grafting reaction on the matrix skeleton of dextran-based crystal gel microspheres, and the reaction time was 2 h to obtain the separation of dextran-based crystal gel microspheres. The medium has a particle size of 500-950 μm, a pore size of 1-20 μm, an effective porosity of 93%, a maximum porosity of 96%, and an ad...

Embodiment 3

[0026] Use 4 mL of an aqueous solution of polymerizable monomers containing 10% of the total concentration of Dex-GMA as the aqueous phase solution; use ethyl acetate as the oil phase solution, at a water phase flow rate of 0.3 cm / s and an oil phase flow rate of 3 cm / s Perform microfluidic focusing into droplets, and form polymerizable droplets with a particle size of 490-2000 μm in 100 mL of ethyl acetate; the polymerizable droplets undergo crystallization and polymerization to obtain dextran-based crystal gel microspheres Matrix skeleton: 30 mL of AMPSA solution with a concentration of 1 M was used to control the temperature at 40-55°C, and the matrix skeleton of dextran-based crystal gel microspheres was grafted for 2 h to obtain dextran-based crystal gel Microsphere separation medium with a particle size of 490-2000 μm, a pore size of 1-30 μm, an effective porosity of 78%, a maximum porosity of 93%, and an adsorption capacity of 10.0 mg / mL crystal gel for lysozyme model pro...

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Abstract

The invention relates to a glucosan-based cryogel microsphere separating medium with superlarge pores for biological separation and a preparation method of the glucosan-based cryogel microsphere separating medium. The glucosan-based cryogel microsphere separating medium is characterized in that supermacroporous glucosan-based cryogel microspheres are porous particles; particles sizes of the supermacroporous glucosan-based cryogel microspheres are 490 to 2000(mu)m, the aperture is 1 to 30(mu)m, and the porosity is 78 to 96 percent; the adsorption capacity to lysoenzyme model protein reaches 5.2 to 10mg / mL cryogel. The invention also provides a method for preparing the cryogel microspheres by using micro-fluidic forming, phase transformation crystallization hole forming and cross linking. The cryogel microsphere separating medium provided by the invention has the advantages of the superlarge pores, good biocompatibility, excellent permeability, high porosity, greater adsorption capacity to protein, good biological safety and the like. The preparation method provided by the invention is simple, and has a broad application prospect in the field of biological separation; particle sizes of the particles are uniform.

Description

technical field [0001] The invention belongs to the technical field of biochemistry, and in particular relates to a dextran-based crystal gel microsphere separation medium and a preparation method thereof. Background technique [0002] Crystal gel chromatography is a new biological separation method developed in recent years. This technology adopts crystal colloid with ultra-large pores with a pore size ranging from several microns to hundreds of microns as the separation medium. It has high porosity and high permeability. Biomacromolecules in the crystal colloid bed column mainly realize rapid adsorption and separation by convective mass transfer. , allowing conventional genetically engineered bacteria cells or cell fragments to pass through the bed without clogging, and can quickly separate target biomacromolecules from fermentation broth or culture fluid containing complex materials at high flow rates. The chromatographic separation method is easy to operate and high in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F251/00C08F220/58B01J20/26B01J20/30
CPCB01J20/264C08F251/00C08F220/585
Inventor 贠军贤高亚伟张颂红沈绍传姚克俭林东强姚善泾
Owner ZHEJIANG UNIV OF TECH
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