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A kind of preparation method of hydrophilic chromatographic medium

A technology of hydrophilic chromatography and chromatographic media, applied in separation methods, chemical instruments and methods, solid adsorbent liquid separation, etc., can solve problems such as low market share, poor quality stability, and few product types

Active Publication Date: 2016-08-24
南通江海港建设工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, some domestic enterprises, research institutes and universities are engaged in the research and development of this medium, but the product types are few, the quality stability is poor, and the market share is very low, which is an urgent strategic problem to be solved in the development of my country's biotechnology industry.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] Take 250ml of dextran gel microspheres with a concentration of 30wt% and an average particle size of 60-100μm, wash them with water and methanol several times on a separatory funnel, transfer them to a reaction bottle or reactor, add 400-600ml of methanol; add 2-8g Glyceride, stir for 30-45min; add 20-35ml of 20wt% ammonia water, stir for 1-2h; add methanol and stir; wash with 2-8 times the volume of methanol for 15-30min, and wash with 1-3 times the volume of water for 10-20min ; Add 200-400ml of water, add 5-15g of potassium carbonate, stir for 2-3h; add 10-30ml of 20% ammonia water, stir for 1-2h; add 10-20ml of chloroform and stir for 6-8h; adjust the pH to 2 with nitric acid -4; add 10-45ml of bromine water, stir for 2-3h; add 20-50g of methacrylic acid, 10-20ml of 20% ammonia, stir for 5-8h; adjust the pH to 2-4 with hydrochloric acid; use 10-20 times the volume Wash with water to obtain a hydrophilic chromatographic medium.

Embodiment 2

[0012] Take 300ml of dextran gel microspheres with a concentration of 30wt% and an average particle size of 60-100μm, wash them with water and methanol several times on a separatory funnel, transfer them to a reaction bottle or reactor, add 400-600ml of methanol; add 2-8g Glyceride, stir for 30-45min; add 20-35ml of 20wt% ammonia water, stir for 1-2h; add methanol and stir; wash with 2-8 times the volume of methanol for 15-30min, and wash with 1-3 times the volume of water for 10-20min ; Add 200-400ml of water, add 5-15g of potassium carbonate, stir for 2-3h; add 10-30ml of 20% ammonia water, stir for 1-2h; add 10-20ml of chloroform and stir for 6-8h; adjust the pH to 2 with nitric acid -4; add 10-45ml of bromine water, stir for 2-3h; add 20-50g of methacrylic acid, 10-20ml of 20% ammonia, stir for 5-8h; adjust the pH to 2-4 with hydrochloric acid; use 10-20 times the volume Wash with water to obtain a hydrophilic chromatographic medium.

Embodiment 3

[0014] Take 350ml of dextran gel microspheres with a concentration of 30wt% and an average particle size of 60-100μm, wash them with water and methanol several times on a separatory funnel, transfer them to a reaction bottle or reactor, add 400-600ml of methanol; add 2-8g Glyceride, stir for 30-45min; add 20-35ml of 20wt% ammonia water, stir for 1-2h; add methanol and stir; wash with 2-8 times the volume of methanol for 15-30min, and wash with 1-3 times the volume of water for 10-20min ; Add 200-400ml of water, add 5-15g of potassium carbonate, stir for 2-3h; add 10-30ml of 20% ammonia water, stir for 1-2h; add 10-20ml of chloroform and stir for 6-8h; adjust the pH to 2 with nitric acid -4; add 10-45ml of bromine water, stir for 2-3h; add 20-50g of methacrylic acid, 10-20ml of 20% ammonia, stir for 5-8h; adjust the pH to 2-4 with hydrochloric acid; use 10-20 times the volume Wash with water to obtain a hydrophilic chromatographic medium.

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PUM

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Abstract

The invention discloses a preparation method of a hydrophilic chromatography medium. The chromatography medium takes methacrylic acid as a ligand and dextrane gel as a substrate. The preparation method of the hydrophilic chromatography medium comprises the following steps: with 200-400ml of dextrane gel microspheres of which the concentration is 30wt% and the average grain diameter is 60-100 microns, washing in a separating funnel by using water and methanol for a plurality of times, and transferring into a reaction bottle or a reactor; adding 400-600ml of methanol, adding 2-8g of glyceride and stirring for 30-45 minutes; adding 20-35ml of 20wt% ammonium hydroxide and stirring for 1-2 hours; adding methanol, stirring, washing by using methanol the volume of which is 2-8 times that of the solution for 15-30 minutes, and washing by water the volume of which is 1-3 times that of the solution for 10-20 minutes; adding 200-400ml of water, adding 5-15g of potassium carbonate, and stirring for 2-3 hours; adding 10-30ml of 20% ammonium hydroxide, and stirring for 1-2 hours; adding 10-20ml of trichloromethane and stirring for 6-8 hours; adjusting the pH to be 2-4 by using nitric acid, adding 10-45ml of bromine water and stirring for 2-3 hours; and adding 20-50g of methacrylic acid and 10-20ml of 20% ammonium hydroxide, stirring for 5-8 hours, adjusting the pH to be 2-4 by using hydrochloric acid, and washing by water the volume of which is 10-20 times that of the solution, thereby obtaining the hydrophilic chromatography medium. The chromatography medium prepared by the preparation method is high in selectivity, and is especially suitable for separation and purification of polyphenol proteins, hydrophilic peptides and natural small organic molecules.

Description

technical field [0001] The invention relates to a preparation method of a hydrophilic chromatographic medium. Background technique [0002] The cost of separation and purification in the production of biopharmaceuticals accounts for the vast majority of the cost, among which the most commonly used is chromatographic technology and its combination. Currently, the media used are mainly gel filtration, ion exchange, hydrophobic interaction, affinity, and reversed phase media. , and its matrix materials include agarose, polystyrene and silica gel. Dextran media is the most widely used matrix material in biopharmaceutical production. It is a natural polysaccharide with high porosity; high connectivity effectively ensures intramolecular substance transfer; large specific surface area provides high adsorption capacity; it is easy to prepare spherical particles of different dextran concentrations and sizes to Meet the needs of different fields; low non-specific adsorption enables ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/291B01J20/30
CPCB01D15/20B01D15/305B01J20/24B01J20/28047
Inventor 华玉叶
Owner 南通江海港建设工程有限公司