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

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Problems solved by technology

The reason is that the complex structure and huge size of macromolecula

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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Example Embodiment

[0020] Example 1

[0021] Take 10g of agarose gel microspheres and use tryptamine as ligand to prepare mixed mode chromatography media. The medium was drained, 3 times the saturated adsorption capacity protein was added, and the adsorption was carried out in 20 mM buffer, pH 5, temperature 25°C, 120 rpm shaker, 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. Wash the medium with 0.1M hydrochloric acid solution, 0.1M sodium hydroxide solution and deionized water in sequence to destroy the hydrophobic interaction 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.

Example Embodiment

[0022] Example 2

[0023] Take 10g of agarose gel microspheres and use tryptamine as ligand to prepare mixed mode chromatography media. The medium was drained, 3 times the saturated adsorption capacity protein was added, and the adsorption was carried out in 20 mM buffer, pH 5, temperature 25°C, 120 rpm shaker, 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. Wash the medium with 0.1M hydrochloric acid solution, 0.1M sodium hydroxide solution and deionized water in sequence to destroy the hydrophobic interaction 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.

Example Embodiment

[0024] Example 3

[0025] Take 10g of agarose gel microspheres and use tryptamine as ligand to prepare mixed mode chromatography media. The medium was drained, 3 times the saturated adsorption capacity protein was added, and the adsorption was carried out in 20 mM buffer, pH 5, temperature 25°C, 120 rpm shaker, 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 0.1 M hydrochloric acid solution, 0.1 M sodium hydroxide solution and deionized water to wash the medium in sequence to destroy the hydrophobic interaction 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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