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High carrying capacity metal chelating affinity chromatography medium

A chromatographic medium and metal chelation technology, applied in chemical instruments and methods, peptide preparation methods, solid adsorbent liquid separation, etc., can solve problems such as single structure, low protein loading, and low separation and purification efficiency, and achieve The improvement method is simple, the protein loading capacity is increased, and the effect of immobilization capacity is improved

Active Publication Date: 2018-03-06
SUZHOU BOJIN BIOLOGICAL TECH
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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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  • High carrying capacity metal chelating affinity chromatography medium

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Embodiment 1

[0016] The preparation process is as follows:

[0017] 1) Soak 20g of silica gel microspheres with a particle size of 30-100μm in 0.5mol / L sodium hydroxide solution, stir at 50-55°C for 1-1.5 hours to activate the silica gel microspheres, and vacuum-dry the silica gel microspheres 12 hours; after taking out the silica gel microspheres from the vacuum drying oven, quickly place them in an anhydrous ethanol solution containing 500g / L carboxymethyl chitooligosaccharide molecules and 3mol / L sodium hydroxide, and stir and react at 35-40°C for 4 ~5 hours, after drying, the inner core of silica gel microspheres with carboxymethyl chitosan molecules attached;

[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 deionized water in sequence, Stir at 50°C for 18 hours, wash with deionized water, and dry to obtain the activated core of silica...

Embodiment 2

[0023] In the step 2) in the embodiment 1, 2mol / L nickel acetate is replaced with the iron acetate of the same amount, all the other preparation processes are consistent with embodiment 1, obtain the bimetallic layer chelation 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 chromatographic medium, in particular to a high-capacity metal chelating affinity chromatographic medium. Background technique [0002] High-efficiency separation and purification of proteins is the premise and basis for the smooth development of biopharmaceuticals, genetic engineering, and life science research. According to the size of the protein itself, the solubility and the number of charges, the methods of purifying proteins are also different. Among them, metal chelation affinity chromatography is based on the separation and purification of proteins based on the different affinity between some amino acid residues on the surface of proteins and immobilized metal ions. The advantages of stability, continuous use or regeneration of the chromatographic column have attracted more and more people's attention. [0003] The metal chelate affinity chromatography medium is mainly composed of three parts: solid matrix, chelatin...

Claims

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Application Information

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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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