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Hydrophobic charge-induced chromatographic medium with bi-functional groups

A technology of hydrophobic charges and bifunctional groups, applied in alkali metal oxides/hydroxides, inorganic chemistry, peptides, etc., can solve the problems of high cost, easy inactivation, binding stability, etc., and achieve convenient cleaning and regeneration, Improved binding with limited selectivity and optimized spatial arrangement

Inactive Publication Date: 2018-05-04
SUZHOU BOJIN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, affinity chromatography uses the reversible combination of target antibodies and specific functional groups to achieve efficient separation and purification of biologically active substances. High cost, easy inactivation, poor binding stability and other disadvantages, new affinity ligands and new adsorption principles have been extensively studied in recent years

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] A method for preparing a hydrophobic charge-induced chromatography medium with dual functional groups, comprising the following steps: taking 10 g of PGMA-EDMA microspheres, adding 2 g of 20% v / v dimethyl sulfoxide, and 2 g of cyanogen bromide and 3g of sodium hydroxide, activated in a 200rpm shaker at 27°C for 30 hours, suction filtered, and washed with deionized water to obtain an activated chromatography matrix; mix the activated chromatography matrix with 2g of N-bromosuccinimide for bromination Alcoholation, reaction in a shaker at 27°C at 200rpm for 2 hours, suction filtration, and washing with deionized water to obtain a bromoalcoholated chromatography matrix; mix the bromoalcoholated chromatography matrix with 4g histidine and 0.5M sodium carbonate buffer Mix, the pH of the sodium carbonate buffer is 10, and react in a 200rpm shaker at 27°C for 6 hours to obtain a histidine-coupled medium; take the histidine-coupled medium and use volume percentages of 20% and 50...

Embodiment 2

[0021] A method for preparing a hydrophobic charge-induced chromatography medium with bifunctional groups, comprising the following steps: taking 10 g of PGMA-EDMA microspheres, adding 3 g of 20% v / v dimethyl sulfoxide, 2 g of brominated Cyanide and 5g sodium hydroxide, activated in a 200rpm shaker at 27°C for 32 hours, suction filtered, and washed with deionized water to obtain an activated chromatography matrix; mix the activated chromatography matrix with 2g N-bromosuccinimide for bromine Alcoholation, reaction in 200rpm shaker at 27°C for 3 hours, suction filtration, washing with deionized water to obtain bromoalcoholation chromatography matrix; buffer the bromoalcoholation matrix with 5g histidine and 0.6M sodium carbonate mixed solution, the pH of the sodium carbonate buffer solution was 10, and reacted in a 200rpm shaker at 27°C for 6 hours to obtain a histidine-coupled medium; take the histidine-coupled medium and use volume percentages of 20%, 50% and % and 80% aceton...

Embodiment 3

[0023] A method for preparing a hydrophobic charge-induced chromatography medium with dual functional groups, comprising the following steps: taking 10 g of PGMA-EDMA microspheres, adding 4 g of 20% v / v dimethyl sulfoxide, and 5 g of cyanogen bromide and 7g sodium hydroxide, activated in a 200rpm shaker at 27°C for 39 hours, suction filtered, and washed with deionized water to obtain an activated chromatography matrix; mix the activated chromatography matrix with 4g N-bromosuccinimide for bromination Alcoholation, react in a shaker at 200 rpm at 27°C for 5 hours, filter with suction, wash with deionized water to obtain a bromoalcoholated chromatography matrix; mix the bromoalcoholated chromatography matrix with 6g histidine and 1M sodium carbonate buffer , the pH of the sodium carbonate buffer solution is 10, reacted in a 200rpm shaker at 27°C for 8 hours to obtain a histidine-coupled medium; take the histidine-coupled medium, use volume percentages of 20%, 50% and Wash with 8...

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Abstract

The invention relates to a hydrophobic charge-induced chromatographic medium with bi-functional groups. The hydrophobic charge-induced chromatographic medium with the bi-functional groups comprises achromatographic matrix and ligands, wherein the chromatographic matrix is made of hydrophilic porous microspheres with hydroxyl groups; the ligands are bifunctional ligands formed after a reaction ofcarboxyl groups of histidine and amino groups of 2-aminobenzothiazole. The novel hydrophobic charge-induced chromatographic medium is prepared by using the histidine and the 2-aminobenzothiazole as the bifunctional ligands, so that the advantages of the conventional hydrophobic charge-induced chromatographic medium are retained; through the introduction of a concept of the bifunctional groups, thespatial arrangement of the ligands is optimized and the antibody bonding selectivity is improved. Therefore, the hydrophobic charge-induced chromatographic medium with the bifunctional groups has a unique separation property.

Description

technical field [0001] The invention relates to a hydrophobic charge-induced chromatographic medium with bifunctional groups, which belongs to the protein chromatographic separation technology in the field of biochemical industry. Background technique [0002] Antibodies can be classified according to different preparation methods: serum polyclonal antibodies obtained by immunizing animals with purified antigens; monoclonal antibodies prepared by hybridoma cell technology; genetically engineered antibodies developed by genetic engineering means, regardless of the preparation method used The obtained antibodies must be isolated and purified. At present, according to the different sources, uses, and characteristics of the antibody itself, if the final requirements can be met, using as few purification steps as possible to increase the yield can effectively improve the purification efficiency. In specific applications, a combination of different separation methods is used to o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/22B01J20/28B01J20/30C07K1/20C07K16/00
CPCB01J20/22B01J20/28021B01J2220/4812C07K1/20C07K16/00
Inventor 瞿欢欢朱至放
Owner SUZHOU BOJIN BIOLOGICAL TECH
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