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Mixing mode expanded adsorbent bed medium and method for producing the same

A technology of mixed mode and adsorption medium, which is applied in the field of expanded bed adsorption and separation of proteins in chromatographic separation, to achieve the effect of large adsorption capacity, high selectivity, and efficient separation and purification

Inactive Publication Date: 2008-10-08
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, the present invention uses cellulose / inorganic weighting agent composite microspheres as the expanded bed adsorption matrix, and the sulfone group introduced after divinyl sulfone activation and the coupled mercaptobenzimidazole are used as its main functional ligands to form a A new type of thiophilic mixed-mode expanded adsorption bed medium, which can be used for large-scale expanded bed adsorption and separation of antibodies, and has become a new type of efficient and integrated antibody separation process. Related research has not been reported in the literature

Method used

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  • Mixing mode expanded adsorbent bed medium and method for producing the same
  • Mixing mode expanded adsorbent bed medium and method for producing the same
  • Mixing mode expanded adsorbent bed medium and method for producing the same

Examples

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Embodiment 1

[0016] Stir 50g of cellulose viscose and 40g of tungsten carbide powder in a 500ml round-bottomed flask at 300rpm, add 300g of pump oil, rotate at 700rpm, inversely suspend and regenerate to make balls, wash with boiling water and deionized water for 3 to 5 times, Screen cellulose / inorganic weighting agent composite microspheres with a particle size of 50-250 μm as the matrix; 10ml of drained composite microspheres, 3.6ml of divinyl sulfone, 9ml of 0.25M pH12 sodium carbonate buffer and 1.5ml of bismuth Methyl sulfoxide was added into the Erlenmeyer flask, activated in a shaker at 180 rpm at 25°C for 4 hours, the mixed solution was removed by suction filtration, washed with deionized water, and sucked dry. Add 2-mercaptobenzimidazole with 3 times the molar amount of double bonds and 10ml of 0.5M sodium hydroxide solution containing 25mg / ml ammonium persulfate, perform coupling reaction in a shaker at 180rpm at 60°C for 8 hours, and remove the mixed solution by suction filtratio...

Embodiment 2

[0018] Stir 50g of cellulose viscose and 40g of tungsten carbide powder in a 500ml round-bottomed flask at 300rpm, add 300g of pump oil, rotate at 700rpm, inversely suspend and regenerate to make balls, wash with boiling water and deionized water for 3 to 5 times, Screen cellulose / inorganic weighting agent composite microspheres with a particle size of 50-250 μm as the matrix; 10ml of drained composite microspheres, 0.9ml of divinyl sulfone, 9ml of 0.5M pH12 sodium carbonate buffer and 1.5ml of bismuth Methyl sulfoxide was added into the Erlenmeyer flask, activated in a shaker at 180 rpm at 25°C for 4 hours, the mixed solution was removed by suction filtration, washed with deionized water, and sucked dry. Add 2-mercaptobenzimidazole with 3 times the molar amount of double bonds and 10ml of 0.7M sodium hydroxide solution containing 25mg / ml ammonium persulfate, perform coupling reaction in a shaker at 180rpm at 60°C for 8 hours, and remove the mixed solution by suction filtration...

Embodiment 3

[0020] Stir 50g of cellulose viscose and 80g of tungsten carbide powder in a 500ml round-bottomed flask at 300rpm, add 310g of pump oil, rotate at 800rpm, inversely suspend and thermally regenerate to make balls, wash with boiling water and deionized water for 3 to 5 times, Screen cellulose / inorganic weighting agent composite microspheres with a particle size of 50-250 μm as the matrix; 10ml of drained composite microspheres, 1.8ml of divinyl sulfone, 9ml of 0.1M pH12 sodium carbonate buffer and 1.5ml of bismuth Methyl sulfoxide was added into the Erlenmeyer flask, activated in a shaker at 180 rpm at 25°C for 4 hours, the mixed solution was removed by suction filtration, washed with deionized water, and sucked dry. Add 2-mercaptobenzimidazole with 3 times the molar amount of double bonds and 10ml of 0.2M sodium hydroxide solution containing 25mg / ml ammonium persulfate, perform coupling reaction in a shaker at 180rpm at 60°C for 8 hours, and remove the mixed solution by suction ...

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Abstract

The invention discloses a mixed model expanded adsorbent bed medium and a preparation method thereof. In the composition of medium, matrix is a composite microsphere with fibrin / inorganic weighting agent, and petunidin is sulfonyl group and mercapto benzimidazole. The composite microsphere with fibrin / inorganic weighting agent is acquired by inverse suspension thermal regeneration and is taken as the matrix to mix divinyl sulfone, soda buffer solution and DMSO for activation, then swab-off activation matrix, the mercapto benzimidazole and sodium hydroxide containing ammonium persulfate are mixed for coupling, thus acquiring the mixed model expanded adsorbent bed medium taking the mercapto benzimidazole and the sulfonyl group as the petunidin. The novel expanded bed adsorbing medium developed by the invention combines with a plurality of petunidin-protein interaction mechanisms such as hydrophobic interaction, charge induction action, thiophilic action, etc, to form mixed model adsorption, which has the advantages of large adsorption capacity, high selectivity, etc., thus being used for high efficient separation and purification of bioactivators such as antibody, etc.

Description

technical field [0001] The invention relates to a mixed-mode expanded bed adsorption medium and a preparation method thereof, which uses mercaptobenzimidazole and sulfone group as ligands, and belongs to the expanded bed adsorption and separation protein technology in chromatographic separation. Background technique [0002] Antibodies are an important biotechnology product widely used in immunotherapy, disease prevention, and medical diagnosis. In recent years, the separation technology for antibodies has made some progress, but it still needs to be continuously supplemented and improved. [0003] Conventional ion-exchange chromatography and hydrophobic interaction chromatography can bind antibodies, but their specificity and selectivity are poor, and there are many separation steps, which affect the activity yield of antibodies. Protein A affinity chromatography media can selectively adsorb antibodies, but the media is expensive, and the ligand is easily degraded by prote...

Claims

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Application Information

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IPC IPC(8): B01J20/22B01J20/30C07K1/22
Inventor 夏海锋林东强姚善泾王立平
Owner ZHEJIANG UNIV
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