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Method for synthesizing hydrophilic chromatographic medium by using trihydroxymethyl aminomethane genin and agarose

A trishydroxymethylaminomethane, synthesis method technology, applied in separation methods, chemical instruments and methods, solid adsorbent liquid separation, etc., can solve the problems of few types and low market share

Inactive Publication Date: 2009-10-21
北京康铭优盛生化技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few types of agarose matrix medium products in my country, and the market share is low. More than 98% of the domestic market is occupied by foreign products. The development of agarose medium with independent intellectual property rights is an urgent strategic problem to be solved.

Method used

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  • Method for synthesizing hydrophilic chromatographic medium by using trihydroxymethyl aminomethane genin and agarose
  • Method for synthesizing hydrophilic chromatographic medium by using trihydroxymethyl aminomethane genin and agarose
  • Method for synthesizing hydrophilic chromatographic medium by using trihydroxymethyl aminomethane genin and agarose

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Embodiment 1

[0035] Embodiment 1, Synthesis Method 1 of Sepharose Hydrophilic Adsorption Chromatography Medium of Tris Hydroxymethylaminomethane

[0036] 1) 100ml of agarose gel microspheres with a concentration of 12% and an average particle size of 30μm are washed with water on a sand core funnel and replaced with acetone, transferred to a reaction bottle or reactor, and 100ml of acetone is added;

[0037] 2) Add 10g of glycidyl ether and stir for 60min;

[0038] 3) Add 3ml of 20% sodium hydroxide and stir for 8h;

[0039] 4) Add acetone and stir for 1h;

[0040] 5) Wash with 5 times the volume of acetone for 10 minutes, and wash with 5 times the volume of water for 10 minutes;

[0041] 6) Add 200ml of water, add 5g of sodium sulfate, and stir for 2h;

[0042] 7) Add 3ml of 20% sodium hydroxide and stir for 1h;

[0043] 8) Add 10ml of epichlorohydrin and stir at 35-60°C for 3h;

[0044] 9) wash with 5 times the volume of water;

[0045] 10) Add 200ml of water, add 5g of sodium sulf...

Embodiment 2

[0055] Embodiment 2, Synthesis Method 2 of Sepharose Hydrophilic Adsorption Chromatography Medium of Tris Hydroxymethylaminomethane

[0056] 1) 100ml of agarose gel microspheres with a concentration of 12% and an average particle size of 30μm are washed with water on a sand core funnel and replaced with acetone, transferred to a reaction bottle or reactor, and 100ml of acetone is added;

[0057] 2) Add 30g glycidyl ether and stir for 120min;

[0058] 3) Add 3ml of 20% sodium hydroxide and stir for 12h;

[0059] 4) add acetone and stir for 2h;

[0060] 5) Wash with 5 times the volume of acetone for 20 minutes, and wash with 5 times the volume of water for 20 minutes;

[0061] 6) Add 200ml of water, add 15g of sodium sulfate, and stir for 3h;

[0062] 7) Add 3ml of 20% sodium hydroxide and stir for 2h;

[0063] 8) Add 10-30ml of epichlorohydrin and stir at 40°C for 5h;

[0064] 9) wash with 5 times the volume of water;

[0065] 10) Add 200ml of water, add 15g of sodium sulfa...

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Abstract

The invention relates to a method for synthesizing a hydrophilic adsorption chromatographic medium by using trihydroxymethyl aminomethane as a genin and agarose gel with high concentration and high crosslinking degree as a matrix. The medium surface of the agarose gel with high concentration and high crosslinking degree has high-density hydroxy for reaction; the agarose gel is activated by allyl bromide, epoxy chloropropane, 1-4-di-butane diglycidyl ether, pentaerythritol tetrol-triglycidyl ether, pentaerythritol tetrol-glycidyl ether or bromhydrin; and then the trihydroxymethyl aminomethane as the genin is added into the agarose gel, and the prepared medium is used as a hydrogen bond donor in a hydrophilic adsorption reaction. The medium prepared by the synthesizing method has the advantages of high rigidity, high genin density and strong selectivity, and is suitable for separating polyphenol protein, hydrophilic polypeptide and natural organic small molecules.

Description

technical field [0001] The invention relates to a method for synthesizing a chromatographic medium, in particular to a method for synthesizing a trishydroxymethylaminomethane agarose hydrophilic chromatographic medium. [0002] The invention belongs to the field of biochemical separation of bioengineering. Background technique [0003] 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, dextran, polystyrene and silica gel. Agarose medium is the most widely used matrix material in the production of biopharmaceuticals. It is a natural polysaccharide with high porosity; high connectivity effectively ensures intramolecular substance transfer; large spec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/30B01J20/22
CPCB01D15/305
Inventor 孟桂凤邢思亮
Owner 北京康铭优盛生化技术有限公司
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