Expanded bed adsorption medium with sulfhydryl ethylpyridine and sulfone group as ligand separation antibody and preparation method

A technology of mercaptoethyl pyridine and adsorption medium, which is applied in the field of expanded bed adsorption and separation of proteins in chromatographic separation, can solve the problems of low antibody selectivity and no emphasis on thiophilic effect.

Inactive Publication Date: 2008-10-15
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, none of these media emphasizes the thiophilic interaction, especially the importance of the sulfone group for antibody binding, so the antibody selectivity is generally not high

Method used

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  • Expanded bed adsorption medium with sulfhydryl ethylpyridine and sulfone group as ligand separation antibody and preparation method
  • Expanded bed adsorption medium with sulfhydryl ethylpyridine and sulfone group as ligand separation antibody and preparation method
  • Expanded bed adsorption medium with sulfhydryl ethylpyridine and sulfone group as ligand separation antibody and preparation method

Examples

Experimental program
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Effect test

Embodiment 1

[0015] 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 4-mercaptoethylpyridine 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

[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, 0.9ml of divinyl sulfone, 9ml of 0.2M 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 4-mercaptoethylpyridine with 3 times the molar amount of double bonds and 10ml of 0.8M 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

[0019] 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 0.5M pH12 sodium carbonate buffer and 1.5ml of di 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 4-mercaptoethylpyridine with 3 times the molar amount of double bonds and 10ml of 0.3M 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 filtr...

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Abstract

The invention discloses an expanded bed adsorption medium for separating antibodies by taking mercapto ehtylpyridine and sulfonyl groups as petunidins, and a preparation method. In the composition of the medium, interstitial substance is cellulose / inorganic weighting-agent composite microspheres, and the petunidins are sulfonyl groups or mercapto ehtylpyridine. The cellulose / inorganic weighting-agent composite microspheres which are used as the interstitial substance are obtained by adopting an opposite-phase suspension thermal-reproduction method; dried activation interstitial substance, the mercapto ehtylpyridine and sodium hydroxide containing ammonium persulfate are mixed for coupling to obtain the expanded bed adsorption medium for separating the antibodies by taking the mercapto ehtylpyridine and the sulfonyl groups as the petunidins. The novel expanded bed adsorption medium developed in the invention enhances the selectivity of mercapto ehtylpyridine petunidin to the antibodies through bring in the sulfonyl groups, has good adsorption separation selectivity to the antibodies, and can be used for the large-scale preparation of the antibodies.

Description

technical field [0001] The invention relates to an expanded bed adsorption medium and a preparation method for separating antibodies with mercaptoethylpyridine and sulfone groups as ligands, and belongs to the expanded bed adsorption and separation protein technology in chromatographic separation. Background technique [0002] Biological downstream technology is an indispensable and important link in the industrialization of bio-industrial technology, and an important guarantee for the yield and quality of biological products. Reasonable design and optimization of downstream processing technology and integration of processes and technologies are crucial to the realization of large-scale biopharmaceuticals. Production is of great significance. [0003] In recent years, new technologies for separation and purification of antibodies have received widespread attention. Antibody products often require high purity and at the same time maintain biological activity as much as possi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
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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