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Preparation method of high-efficiency hydrophobic medium and application of high-efficiency hydrophobic medium in purification of defibrase

A technology of hydrophobic medium and defibrase, which is applied in the field of biochemistry and achieves the effects of high yield, simple operation and high purity

Active Publication Date: 2013-06-12
江西浩然生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] 4. No non-specific adsorption: the matrix itself will not adsorb proteins under chromatographic operating conditions

Method used

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  • Preparation method of high-efficiency hydrophobic medium and application of high-efficiency hydrophobic medium in purification of defibrase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1) Weigh 1g of BPO, 5g of PVP, 50g of GMA, 600ml of ethanol, and 70ml of toluene. Add the weighed raw materials into an appropriate glass reaction bottle, make it fully dissolved and blow nitrogen to remove oxygen. Then place the reaction bottle on a device with a rotating and heating device, and polymerize at 75 degrees Celsius for 24 hours. After the reaction, the solvent was removed by sedimentation, washed with ethanol, and dried in vacuum for 24 hours to obtain monodisperse PGMA microspheres.

[0023] 2) Put 50g of PGMA microspheres obtained in the first step into a 250ml glass reactor (with a water bath), add 50ml of dioxane, adjust the temperature of the water bath to 50°C, and add 0.5 ml, stirred for 5 minutes, then added 5ml of n-butanol and continued to stir, the reaction time was 30 minutes, collected microspheres, washed with purified water and dioxane alternately three times each, and finally washed twice with dioxane; dried to obtain butanol Base polymer...

Embodiment 2

[0026] 1) Weigh 15g of BPO, 100g of PVP, 750g of GMA, 7000ml of ethanol, and 1200ml of toluene. Add the weighed raw materials into a 20L glass reaction bottle, make it fully dissolved and blow nitrogen to remove oxygen. Then place the reaction bottle on a device with a rotating and heating device, and polymerize at 70 degrees Celsius for 30 hours. After the reaction, the solvent was removed by sedimentation, washed with ethanol, and dried in vacuum for 24 hours to obtain monodisperse PGMA microspheres.

[0027] 2) Put 375g of PGMA microspheres obtained in the first step into a 1000ml glass reactor (with a water bath), add 375ml of dioxane, adjust the temperature of the water bath to 45°C, add in the catalyst (boron trifluoride ether) 3.75 ml, after stirring for 5 minutes, add 40ml of n-octanol and continue stirring, the reaction time is 40min, collect microspheres, wash with purified water and dioxane alternately three times each, and finally wash twice with dioxane; dry to o...

Embodiment 3

[0030]1) Weigh 15g of AIBN, 120g of PVP, 650g of GMA, 6500ml of ethanol, and 1000ml of toluene. Add the weighed raw materials into a 20L glass reaction bottle, make it fully dissolved and blow nitrogen to remove oxygen. Then place the reaction bottle on a device with a rotating and heating device, and polymerize at 75 degrees Celsius for 36 hours. After the reaction, the solvent was removed by sedimentation, washed with ethanol, and dried in vacuum for 24 hours to obtain monodisperse PGMA microspheres.

[0031] 2) Weigh 375g of PGMA microspheres obtained in the first step, put them in a 1000ml glass reactor (with a water bath), add 375ml of dioxane, adjust the temperature of the water bath to 50°C, and add in the catalyst (boron trifluoride ether) 4ml, stirred for 5min, added 40ml of liquid phenol and continued to stir, the reaction time was 50min, collected microspheres, washed with purified water and dioxane alternately three times each, and finally washed twice with dioxan...

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Abstract

The invention discloses a preparation method of a high-efficiency hydrophobic medium and an application of the high-efficiency hydrophobic medium in purification of defibrase. The preparation method comprises the following steps of: 1) preparing polyglycidyl methacrylate (PGMA) microspheres by virtue of a dispersion polymerization method; and 2) preparing the high-efficiency hydrophobic medium by taking the polyglycidyl methacrylate (PGMA) microspheres as a matrix, wherein the high-efficiency hydrophobic medium is filled in a stainless steel chromatographic column with a pressure of 20 MPa, and then the column is connected with a liquid chromatography system and used for purifying defibrase. The preparation method disclosed by the invention has the advantages that product purity is high, yield is high, operation is simple and convenient, industrialization requirements can be achieved, and popularization is facilitated.

Description

technical field [0001] The invention belongs to the field of biochemistry, and in particular relates to a preparation method of a high-efficiency hydrophobic medium and its application in purifying defibrase from snake venom. technical background [0002] The matrix of traditional hydrophobic media is usually a soft matrix such as agarose and cellulose, which have the advantages of good biocompatibility, good chemical stability, good permeability, and less non-specific adsorption. However, their poor mechanical properties prevent them from being used in modern high-efficiency preparative liquid phase systems. The silica gel matrix developed in recent years is a hard matrix. The hydrophobic medium synthesized based on it is used with modern high-efficiency preparative liquid chromatography, which has achieved unprecedented improvement in hydrophobic chromatography column efficiency and greatly improved purification efficiency. However, due to the poor chemical stability of s...

Claims

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Application Information

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IPC IPC(8): C08F120/32C08F8/00B01J20/26B01J20/281B01J20/30
Inventor 胡小光刘庆华张素芳
Owner 江西浩然生物制药有限公司
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