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Dual-mode identification chromatographic medium and preparation method thereof

A chromatographic medium and dual-mode technology, applied in chemical instruments and methods, other chemical processes, alkali metal oxides/hydroxides, etc., can solve problems such as mass transfer efficiency and complex size that hinder the imprinting process, and achieve excellent durability Salt adsorption capacity, retention of adsorption performance, effect of preventing competitive adsorption

Active Publication Date: 2018-02-27
TAIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reason is that the complex structure and huge size of macromolecular substances hinder the imprinting process and mass transfer efficiency.

Method used

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  • Dual-mode identification chromatographic medium and preparation method thereof
  • Dual-mode identification chromatographic medium and preparation method thereof
  • Dual-mode identification chromatographic medium and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Get 10g of agarose gel microspheres, and tryptamine is used as a ligand to prepare a mixed-mode chromatography medium. The medium was drained, and protein with 3 times the saturated adsorption capacity was added, and then adsorbed in 20mM buffer solution, the pH was 5, the temperature was 25°C, and the shaker was 120rpm for 12 hours. Add 1:3 tetramethoxysilane and phenyltriethoxysilane to the adsorption system, adjust the pH to 9.3 with ammonia water, and carry out the polymerization reaction. The reaction time is 1 hour. Use 0.1M hydrochloric acid solution, 0.1M sodium hydroxide solution and deionized water to wash the medium in order to destroy the hydrophobic force between the protein and the ligand, desorb the template protein, and obtain a dual-mode recognition chromatography medium. According to the nitrogen adsorption method, the average pore size of the medium is 33.3nm.

Embodiment 2

[0023] Get 10g of agarose gel microspheres, and tryptamine is used as a ligand to prepare a mixed-mode chromatography medium. The medium was drained, and protein with 3 times the saturated adsorption capacity was added, and then adsorbed in 20mM buffer solution, the pH was 5, the temperature was 25°C, and the shaker was 120rpm for 12 hours. Add 1:3 tetramethoxysilane and phenyltriethoxysilane to the adsorption system, adjust the pH to 9.3 with ammonia water, and carry out the polymerization reaction. The reaction time is 2 hours. Use 0.1M hydrochloric acid solution, 0.1M sodium hydroxide solution and deionized water to wash the medium in order to destroy the hydrophobic force between the protein and the ligand, desorb the template protein, and obtain a dual-mode recognition chromatography medium. According to the nitrogen adsorption method, the average pore size of the medium is 27.7nm.

Embodiment 3

[0025] Get 10g of agarose gel microspheres, and tryptamine is used as a ligand to prepare a mixed-mode chromatography medium. The medium was drained, and protein with 3 times the saturated adsorption capacity was added, and then adsorbed in 20mM buffer solution, the pH was 5, the temperature was 25°C, and the shaker was 120rpm for 12 hours. Add 1:3 tetramethoxysilane and phenyltriethoxysilane to the adsorption system, adjust the pH to 9.3 with ammonia water, and carry out the polymerization reaction. The reaction time is 4 hours. Use 0.1M hydrochloric acid solution, 0.1 M sodium hydroxide solution and deionized water to wash the medium sequentially, destroy the hydrophobic force between the protein and the ligand, desorb the template protein, and obtain a dual-mode recognition chromatography medium. According to the nitrogen adsorption method, the average pore size of the medium is 26.7nm.

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Abstract

The invention discloses a dual-mode identification chromatographic medium with a mixed mode and a molecular engram mode and a preparation method thereof. The preparation method comprises the followingsteps of: selecting a mixed mode medium and template protein for full adsorption; carrying out polymerization on the surface of the medium by using tetramethylsilane and phenyl triethoxysilane to form an engram layer; washing the engram medium with a hydrochloric acid solution, a sodium hydroxide solution and deionized water in sequence to remove the template protein so as to obtain the dual-modechromatography medium. The novel chromatographic medium developed by the invention has two identification capabilities of a mixed mode and a molecular engram mode, greatly improves the removal capability of the mixed mode medium to macromolecular impurities by utilizing the steric hindrance of a molecular engram polymer to macromolecular substances, improves the selectivity of chromatography, andretains good adsorption performance, pH dependence and salt-tolerant adsorption capability of the mixed mode.

Description

technical field [0001] The invention relates to a dual-mode recognition chromatographic medium with a mixed mode and a molecular imprinted mode and a preparation method thereof, belonging to protein chromatography separation technology in the field of biochemical industry. Background technique [0002] In recent years, the development of biomacromolecular drugs represented by protein drugs has become more and more rapid, and its application in the field of therapeutic diagnosis has continued to expand, and the proportion of its output value in the entire pharmaceutical field has also continued to increase. Compared with chemical drugs, biological drugs not only require high product purity, but also maintain biological activity, so separation and purification are more difficult. For the entire biopharmaceutical production process, the cost of separation and purification steps is as high as 50-80%, which has become a bottleneck hindering the further industrialization of biopha...

Claims

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Application Information

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IPC IPC(8): B01J20/26B01J20/28B01J20/30C08G77/04C08G77/06
CPCB01J20/268B01J20/28083C08G77/04C08G77/06
Inventor 施伟
Owner TAIZHOU UNIV
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