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A high-capacity metal chelate affinity chromatography medium

A metal chelation and chromatography medium technology, applied in chemical instruments and methods, other chemical processes, separation methods, etc., can solve the problems of single structure, low protein loading, low separation and purification efficiency, etc. Increase protein load and improve the effect of immobilization

Active Publication Date: 2020-07-14
SUZHOU BOJIN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, the existing metal chelate affinity chromatography medium has a single structure, generally has the problems of low protein loading and low separation and purification efficiency

Method used

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  • A high-capacity metal chelate affinity chromatography medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] The preparation process is as follows:

[0017] 1) Soak 20 g of silica gel microspheres with a particle size of 30 to 100 μm in 0.5 mol / L sodium hydroxide solution, stir at 50 to 55 ° C for 1 to 1.5 hours to activate the silica gel microspheres, and vacuum dry the silica gel microspheres 12 hours; after the silica gel microspheres were taken out from the vacuum drying oven, they were quickly placed in anhydrous ethanol solution containing 500g / L carboxymethyl chitosan oligosaccharide molecules and 3mol / L sodium hydroxide, and the reaction was stirred at 35-40°C for 4 ~5 hours, after drying, the inner core of silica gel microspheres with carboxymethyl chitosan oligosaccharide molecules is obtained;

[0018] 2) Rinse the inner core of the silica gel microspheres obtained in step 1) with deionized water, add 23g (about 0.2mol) allyl glycidyl ether, 5g sodium sulfate, 5g sodium hydroxide, and 100ml Stir at 50°C for 18 hours, wash with deionized water, and dry to obtain an ...

Embodiment 2

[0023] In step 2) in Example 1, 2 mol / L of nickel acetate was replaced with the same amount of ferric acetate, and the rest of the preparation process was the same as in Example 1, to obtain a bimetallic layer chelated affinity chromatography medium of different metals.

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Abstract

The invention relates to a high carrying capacity metal chelating affinity chromatography medium. Silica gel microspheres are adopted as a core; pores are formed inside the silica gel microspheres; carboxymethyl chitosan oligosaccharide molecules are adhered to the surfaces and the pores of the silica gel microspheres; a part of the carboxymethyl chitosan oligosaccharide molecules are complexed with first metal ions to form a first layer of metal chelating; the rest part of the carboxymethyl chitosan oligosaccharide molecules are bonded with allyl glycidyl ether; the allyl glycidyl ether is connected with aglycone; and the aglycone is complexed with second metal ions to form a second layer of metal chelating. By adopting the medium, double metal layers are successfully immobilized on the surface of the medium, the carrying capacity of the metal affinity chromatography medium is effectively increased, and the medium is simple in improvement method, low in production cost and beneficialto large-scale popularization and application.

Description

technical field [0001] The invention relates to a chromatography medium, in particular to a high-load metal chelate affinity chromatography medium. Background technique [0002] The efficient separation and purification of proteins is the premise and basis for the smooth development of biopharmaceuticals, genetic engineering, and life science research. Depending on the size of the protein itself, the solubility and the number of charges, the methods of protein purification are also different. Among them, metal chelation affinity chromatography technology is based on the different affinity of some amino acid residues on the protein surface and immobilized metal ions to separate and purify proteins. The advantages of stability, continuous use or regeneration of chromatography columns have attracted increasing attention. [0003] The metal chelate affinity chromatography medium is mainly composed of three parts: solid matrix, chelating ligand and metal ions, and these componen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/24B01J20/30B01D15/38C07K1/22
CPCB01D15/3804B01J20/103B01J20/24B01J2220/4806B01J2220/4812B01J2220/4825C07K1/22
Inventor 瞿欢欢朱至放
Owner SUZHOU BOJIN BIOLOGICAL TECH
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