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Preparation method of hydrophobic charge-induced chromatography medium with bifunctional groups

A technology of hydrophobic charges and bifunctional groups, which is applied in the preparation of hydrophobic charge-induced chromatography media and in the field of protein chromatography separation, can solve problems such as difficult online cleaning, low elution pH, and limited processing capacity, and achieve optimal Spatial arrangement, convenient cleaning and regeneration, and the effect of improving selectivity

Active Publication Date: 2020-06-19
SUZHOU BOJIN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Protein A affinity chromatography is a typical representative, with good selectivity, high resolution, and high-purity antibody products can be obtained by one-step separation, but there are also some limitations, such as high cost, limited processing capacity, and low elution pH , online cleaning difficulties, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Take 10 g of coarsened silicon dioxide microspheres, add 5 g of 20% v / v dioxane, 4 g of diethylene glycol bisglycidyl ether and 3 g of sodium hydroxide, and stir at 40 ° C for 12 hours , filtered with suction, washed with deionized water to obtain an activated chromatography matrix; the activated chromatography matrix was mixed with 3 g of 3-pyrrole carboxylic acid and 0.1-0.5 mol of sodium hydroxide solution, stirred and reacted at 50 ° C for 8-16 hours, Obtain the medium of 3-pyrrole carboxylic acid coupling; take the medium of 3-pyrrole carboxylic acid coupling, wash with ethanol solution with volume percentage of 30%, 70% and 100% respectively, then wash with dimethylformamide, add 10 g of dimethylformamide, 2-amino-1,3,5-triazine and N,N-dicyclohexylcarboimide, 2-amino-1,3,5-2-amino-1,3 , the concentration of 5-triazine is 0.1g / ml, the concentration of N,N-dicyclohexylcarboimide is 0.6g / ml, shake at 50°C for 12 hours to obtain 2-amino-1,3,5-2-amino - 1,3,5-triazin...

Embodiment 2

[0022] Take 10 g of coarsened silicon dioxide microspheres, add 8 g of 20% v / v dioxane, 10 g of diethylene glycol bisglycidyl ether and 4 g of sodium hydroxide, and stir at 40 ° C for 10 hours , filtered with suction, and washed with deionized water to obtain an activated chromatography matrix; the activated chromatography matrix was mixed with 3 g of 3-pyrrole carboxylic acid and 2 g of sodium hydroxide solution, and stirred and reacted at 50 ° C for 10 hours to obtain 3-pyrrole Carboxylic acid coupling medium; take the medium of 3-pyrrole carboxylic acid coupling, wash with 30%, 70% and 100% ethanol solution respectively by volume percentage, then wash with dimethylformamide, add 10g of dimethylformamide Nylformamide, 2-amino-1,3,5-2-amino-1,3,5-triazine and N,N-dicyclohexylcarbimide, 2-amino-1,3,5-2 -Amino-1,3,5-triazine at a concentration of 0.2 g / ml, N,N-dicyclohexylcarboimide at a concentration of 0.8 g / ml, shaking at 50°C for 17 hours to obtain 2-amino-1,3 , 5-2-amino-...

Embodiment 3

[0024] Take 10 g of coarsened silicon dioxide microspheres, add 12 g of 20% v / v dioxane, 15 g of diethylene glycol bisglycidyl ether and 6 g of sodium hydroxide, and stir at 40 ° C for 16 hours , filtered with suction, and washed with deionized water to obtain an activated chromatography matrix; the activated chromatography matrix was mixed with 3 g of 3-pyrrole carboxylic acid and 5 g of sodium hydroxide solution, and stirred and reacted at 50 ° C for 13 hours to obtain 3-pyrrole Carboxylic acid coupling medium; take the medium of 3-pyrrole carboxylic acid coupling, wash with ethanol solution with volume percentage of 30%, 70% and 100%, and then wash with dimethylformamide, add 10g of dimethylformamide Nylformamide, 2-amino-1,3,5-2-amino-1,3,5-triazine and N,N-dicyclohexylcarbimide, 2-amino-1,3,5-2 The concentration of -amino-1,3,5-triazine is 0.5g / ml, the concentration of N,N-dicyclohexylcarboimide is 0.6g / ml, shaken at 50℃ for 20 hours to obtain 2-amino-1,3 , the medium ac...

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PUM

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Abstract

The invention relates to hydrophobic charge-induction chromatography media with double functional groups. The hydrophobic charge-induction chromatography media comprise a chromatography substrate andligands, wherein the chromatography substrate is hydrophilic porous microspheres with hydroxy groups, and the ligands are difunctional ligands formed after reaction of carboxyl groups of pyrrole-3-carboxylic acid and amino groups of 4-amino-2-carbonyl pyrimidine. According to the novel hydrophobic charge-induction chromatography media prepared from the difunctional ligands formed by pyrrole-3-carboxylic acid and 4-amino-2-carbonyl pyrimidine, the concept of the double functional groups is introduced, spatial arrangement of the ligands is optimized, antibody fixation selectivity is improved, and the hydrophobic charge-induction chromatography media have specific separation performance accordingly.

Description

technical field [0001] The invention relates to a preparation method of 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] Antibody (antibody) is one of the most important immune substances in the organism, which participates in the immune process of the body. Due to the high affinity and specificity of antibodies and antigens, they are widely used in the field of clinical medicine. Monoclonal antibody (referred to as monoclonal antibody, monoclonal antibody, mAb), as an important class of antibody drugs, has the advantages of strong specificity, high affinity, and low toxicity and side effects, and is used for the treatment of major diseases such as cancer and immunodiagnosis. Separation and purification is an important part of antibody production and preparation, and the downstream process accounts for about 80% ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/22B01J20/28B01J20/30B01D15/38C07K1/16
CPCB01D15/3885B01J20/22B01J20/28011C07K1/16
Inventor 瞿欢欢朱至放
Owner SUZHOU BOJIN BIOLOGICAL TECH
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