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A metal chelate affinity chromatography medium based on magnetic polymer microspheres

A technology of metal chelation and chromatography media, which is applied in the field of metal chelation affinity chromatography media, can solve the problems of separating proteins that are not suitable for high throughput, limited number of reuses, and low mechanical strength of the media, and achieve good biological Compatibility, long service life, weak swelling effect

Active Publication Date: 2020-06-30
SUZHOU BOJIN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The immobilized metal chelate affinity chromatography is based on the affinity between the target protein and metal ions. The metal ions in the existing agarose gel-based metal chelate affinity chromatography medium The content is low, not suitable for high-throughput protein isolation, and the medium has low mechanical strength, expensive, limited number of reuses, and short life

Method used

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  • A metal chelate affinity chromatography medium based on magnetic polymer microspheres

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The preparation process is as follows:

[0023] (1) Dissolve 18g of sodium alginate, 5g of polyacrylamide, and 2g of silicic acid into 200mL of dilute hydrochloric acid solution with a pH of 3, and stir fully at room temperature to form mixed solution A; Add ferric oxide magnetic balls into 20mL deionized water, and stir vigorously at room temperature to form suspension B;

[0024] (2) Under stirring conditions, slowly add the suspension B into the mixed solution A, and then vigorously stir for 15-20 minutes to obtain the mixed solution C; slowly stir the mixed solution C and raise the temperature to 80-90°C for 1.5-2 hours , and then ultrasonically dispersed to obtain a dispersion, which was then centrifuged to remove large particles;

[0025] (3) Add the dispersion into the syringe, fix the syringe on the syringe pump, add 5kV static electricity to the needle of the syringe, and then squeeze the mixed solution into the 0.1mol / L NaOH solution at a rate of 200mL / h. Ob...

Embodiment 2

[0031] The preparation process is as follows:

[0032] Nickel acetate solution is replaced with the copper acetate solution of identical concentration in the step (7) of embodiment 1, and all the other operating processes are identical with embodiment 1.

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Abstract

The invention relates to a metal chelated affinity chromatography medium with a magnetic polymer microsphere as a matrix. A polymer composite microsphere wrapped with a ferroferric oxide microsphere is adopted as a core, and the polymer is composed of sodium alginate, polyacrylamide and silicic acid; and the surface of the core is cross-linked with chitosan oligosaccharide and cellulose, the chitosan oligosaccharide and cellulose are bonded with allyl glycidyl ether, the allyl glycidyl ether is connected with a ligand, and the ligand is used to complex metal ions. The metal chelated affinity chromatography medium prepared in the invention can be repeatedly used for many times, has strong mechanical properties, can separately complex different metal ions, has a long service life, and has ahigh efficiency in separating target proteins.

Description

technical field [0001] The invention relates to a chromatographic medium, in particular to a metal chelate affinity chromatographic medium with magnetic polymer microspheres as the matrix. Background technique [0002] As one of the most important biological macromolecules, protein is the main embodiment and material basis of all life activities. The precise research on protein structure and its biological function must be based on obtaining high-purity target protein. Protein separation and purification is the The method of separating and purifying the desired target protein from the mixture with the downstream technology of bioengineering is one of the core technologies in the contemporary bioindustry. [0003] The basis for protein separation and purification usually includes: the use of solubility differences, molecular size differences, surface charge differences, hydrophilic and hydrophobic characteristics differences, specific biological affinity differences, etc. Am...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/26B01J20/28B01J20/30B01D15/38
CPCB01D15/3804B01J20/06B01J20/262B01J20/28009B01J2220/4825
Inventor 瞿欢欢朱至放
Owner SUZHOU BOJIN BIOLOGICAL TECH