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High-stability immobilized metal-chelating affinity chromatography medium

A technology for immobilizing metals and chromatographic media, applied in the fields of alkali metal compounds, chemical instruments and methods, alkali metal oxides/hydroxides, etc., which can solve the problems of difficult maintenance of purification and separation effects, poor chemical stability, and the use of media Short service life and other problems, to achieve the effect of increasing stability, increasing mechanical properties, and high media solid loading capacity

Active Publication Date: 2018-03-06
SUZHOU BOJIN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The currently commercialized immobilized metal chelate affinity chromatography media include Ni-NTA Agarose and Ni-IDA Agarose, two kinds of agarose gels that coordinate and bind nickel ions. Due to the preparation process, they are expensive and have poor chemical stability. Good, the service life of the medium is short, and it is difficult to maintain the initial purification and separation effect after multiple washings

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The preparation process is as follows:

[0018] 1) Soak 20 g of agarose gel microspheres with a particle size of 30-100 μm in 0.5 L of deionized water, and stir magnetically at 45-50 ° C for 15 minutes;

[0019] 2) Then add 1.5g of silicon nitride, 1.5g of boron nitride, and 6g of amino oligosaccharin to the system obtained in step 1), stir magnetically at 50-55°C for 1.5-2 hours, cool to room temperature, filter, wash and dry ;

[0020] 3) Place the microspheres obtained in step 2) in 0.5L of sulfuric acid solution with a concentration of 50wt%, and add 5g of allyl-2,3-epoxypropyl ether and 2.5g of sodium sulfate at the same time. Stir for 15 hours, filter and vacuum dry after the reaction;

[0021] 4) Add 15mL of aminotriacetic acid, 60mL of ethanol and 5g of LiOH to the microspheres obtained in step 3), stir and react at 55-60°C for 6-8h, filter to remove the solvent after the reaction, wash and dry;

[0022] 5) Soak the microspheres obtained in step 4) in 1 mol / L...

Embodiment 2

[0024] The preparation process is as follows:

[0025] 1) Soak 20 g of agarose gel microspheres with a particle size of 30-100 μm in 0.5 L of deionized water, and stir magnetically at 45-50 ° C for 15 minutes;

[0026] 2) Then add 1g of silicon nitride, 1g of boron nitride, and 4.5g of amino oligosaccharin to the system obtained in step 1), stir magnetically at 50-55°C for 1.5-2 hours, cool to room temperature, filter, wash and dry;

[0027] 3) Place the microspheres obtained in step 2) in 0.5L of sulfuric acid solution with a concentration of 50wt%, and add 5g of allyl-2,3-epoxypropyl ether and 2.5g of sodium sulfate at the same time. Stir for 15 hours, filter and vacuum dry after the reaction;

[0028] 4) Add 15mL of aminotriacetic acid, 60mL of ethanol and 5g of LiOH to the microspheres obtained in step 3), stir and react at 55-60°C for 6-8h, filter to remove the solvent after the reaction, wash and dry;

[0029] 5) Soak the microspheres obtained in step 4) in 1 mol / L nic...

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Abstract

The invention relates to a high-stability immobilized metal-chelating affinity chromatography medium which adopts agar agarose gel microspheres as an inner core, wherein silicon nitride and boron nitride are dispersed on the surfaces of the agar agarose gel microspheres; oligosaccharins are cross-linked on the surfaces of the agar agarose gel microspheres; hydroxyl in agar agarose molecules on thesurfaces of the agar agarose gel microspheres and amino in the oligosaccharins are bonded with allyl-2,3-allyl glycidyl ether; the allyl-2,3-allyl glycidyl ether is connected with aglucon; and the aglucon is used for immobilizing and loading metal ions through coordination. Through design and improvement of the chromatography medium, the stability of the chromatography medium is improved, the service life of the chromatography medium is prolonged, and a relatively high medium immobilization capacity and a good separation effect can be still maintained after multiple times of repeated use.

Description

technical field [0001] The invention relates to a chromatographic medium, in particular to a highly stable immobilized metal chelating affinity chromatographic medium. Background technique [0002] The separation and purification of proteins has important applications in the fields of genetic engineering, pharmaceutical engineering and other disciplines. Currently, protein separation and purification technologies include: precipitation methods, such as salting out, heavy metal salt precipitation, heating denaturation, alkaloid or acid precipitation, etc.; chromatography, Such as affinity chromatography, adsorption chromatography, gel filtration chromatography, ion exchange chromatography, etc.; electrochemical methods, such as isoelectric focusing, electrophoresis; centrifugation; membrane separation technology, etc. [0003] Among them, the transition metal ions contained in the chromatography medium in the immobilized metal chelate affinity chromatography technology, such ...

Claims

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Application Information

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IPC IPC(8): B01J20/24B01J20/28B01J20/30B01D15/38
CPCB01D15/3804B01J20/0248B01J20/0251B01J20/0259B01J20/24B01J20/28021B01J20/28047B01J2220/4806B01J2220/4812B01J2220/4825
Inventor 瞿欢欢朱至放
Owner SUZHOU BOJIN BIOLOGICAL TECH
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