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Hydrophobic electric charge induced type thiophil expanded adsorbent bed medium and method for producing the same

A technology of hydrophobic charge and adsorption medium, which is applied in the field of expanded bed adsorption and separation of proteins in chromatographic separation. It can solve the problems that the expansion bed adsorption process cannot be used, and there is no clear combination of thiophile, so as to ensure the coupling efficiency and improve the activation. Efficiency, effect of high protein adsorption capacity

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

AI Technical Summary

Problems solved by technology

However, none of these HCIC media clearly binds to thiophile, and is a medium with a density close to 1. It can only be used in conventional packed chromatography, and cannot be used in the newly emerging expanded bed adsorption process.

Method used

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  • Hydrophobic electric charge induced type thiophil expanded adsorbent bed medium and method for producing the same
  • Hydrophobic electric charge induced type thiophil expanded adsorbent bed medium and method for producing the same
  • Hydrophobic electric charge induced type thiophil expanded adsorbent bed medium and method for producing the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] 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-mercapto-1-methylimidazole 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 by suction filtration The mixture...

Embodiment 2

[0019] 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 0.1M 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-mercapto-1-methylimidazole with 3 times the molar amount of double bonds and 10ml of 0.4M sodium hydroxide solution containing 25mg / ml ammonium persulfate, perform coupling reaction in a shaking table at 180rpm at 60°C for 8 hours, and remove by suction filtration The m...

Embodiment 3

[0021] 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 0.4M 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-mercapto-1-methylimidazole with 3 times the molar amount of double bonds and 10ml of 0.6M sodium hydroxide solution containing 25mg / ml ammonium persulfate, conduct coupling reaction in a shaker at 180rpm at 60°C for 8 hours, and remove by suction filtration Th...

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Abstract

The invention discloses a charge-induced sulphophile expanded adsorbent bed medium with hydrophobic nature and a preparation method thereof. In the composition of the medium, matrix is a composite microsphere with fibrin / inorganic weighting agent, 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 active matrix, the mercapto benzimidazole and sodium hydroxide containing ammonium persulfate are mixed for coupling, thus acquiring the charge-induced sulphophile expanded adsorbent bed medium with the hydrophobic nature. The expanded bed adsorbing medium developed by the invention combines with three protein separation principles of hydrophobic charge induced chromatography, thiophilic chromatography and expanded bed adsorption, etc., which has the advantages of large density of the petunidin, large protein adsorption capacity, strong specificity, mild conditions of adsorption and elution, etc., and shows the characteristic of the integration of technology and process, thus being used for high efficient separation and purification of proteins such as antibody, etc.

Description

technical field [0001] The invention relates to a hydrophobic charge-induced thiophilic expanded bed adsorption medium and a preparation method thereof, belonging to the expanded bed adsorption and separation protein technology in chromatographic separation. Background technique [0002] With the rapid development of genetic engineering, protein engineering, fermentation engineering, and cell engineering, more and more biological products require efficient biological downstream technologies for separation and purification, and new requirements have been put forward for biological separation methods. [0003] Antibodies are an important biotechnology product widely used in immunotherapy, disease prevention, and medical diagnosis. The technology of preparing polyclonal antibodies and monoclonal antibodies through cell engineering and immune methods has become more and more mature, the types of antibodies are increasing, and the scale of preparation is also expanding, which req...

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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