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Chromatographic medium with indole-3-acethydrazide as functional ligand

A chromatographic medium, acetyl hydrazide technology, applied in other chemical processes, chemical instruments and methods, etc., can solve the problems of the separation process being difficult to meet the requirements, poor specificity and selectivity, limiting large-scale applications, etc., and achieve a significant hydrophobic charge. Induction chromatography separation characteristics, high coupling efficiency, strong charge induction effect

Inactive Publication Date: 2018-04-27
SUZHOU BOJIN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Antibody products often require high purity and must maintain biological activity, so traditional separation processes are often difficult to meet the requirements
Protein A or protein G affinity chromatography media can specifically bind antibodies, but protein affinity ligands are easily degraded by proteases in the feed solution, contaminating the product, and the number of reuses is low. The media is expensive and the operating cost is extremely high. limit its large-scale application
Although traditional methods such as ion exchange chromatography and hydrophobic interaction chromatography can bind antibodies, they have poor specificity and selectivity, many separation steps, and limited purification effects.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Take 8g of drained agarose gel 4FF, 2g of polyacrylamide, add 5g of 40% (v / v) dioxane, 4g of 1,4-butanediol bisglycidyl ether activator and 6g of hydroxide Sodium, stirred and reacted at 60°C for 10 hours, suction filtered, and washed with deionized water to obtain a chromatographic matrix activated by 1,4-butanediol bisglycidyl ether; the activated layer of 1,4-butanediol bisglycidyl ether The analysis matrix was mixed with 4g of indole-3-acetylhydrazide and 1g of sodium hydroxide, stirred and reacted at 60°C for 20 hours, filtered with suction, washed with deionized water, and obtained with indole-3-acetylhydrazide as the functional ligand Hydrophobic charge-induced chromatography medium with a ligand density of 100 μmol / ml.

Embodiment 2

[0022] Take 8g of drained agarose gel 4FF, 2g of polyacrylamide, add 6g of 40% (v / v) dioxane, 5g of 1,4-butanediol bisglycidyl ether activator and 2g of hydroxide Sodium, stirred and reacted at 60°C for 12 hours, suction filtered, and washed with deionized water to obtain an epoxy-activated chromatography matrix; the epoxy-activated chromatography matrix and 2g of indole-3-acetylhydrazide and 3g of sodium hydroxide Mix, stir and react at 60°C for 12 hours, filter with suction, and wash with deionized water to obtain a hydrophobic charge-induced chromatography medium with indole-3-acetylhydrazide as a functional ligand, with a ligand density of 90 μmol / ml.

Embodiment 3

[0024] Take 8g of drained agarose gel 4FF, 2g of polyacrylamide, add 8g of 40% (v / v) dioxane, 8g of epoxy activator and 6g of sodium hydroxide, stir and react at 60°C for 20 hours , filtered with suction, and washed with deionized water to obtain an epoxy-activated chromatography matrix; mix the epoxy-activated chromatography matrix with 6 g of indole-3-acetylhydrazide and 5 g of sodium hydroxide, and stir and react at 60 ° C for 35 hours , filtered with suction, and washed with deionized water to obtain a hydrophobic charge-induced chromatography medium with indole-3-acetylhydrazide as a functional ligand, and the ligand density is 130 μmol / ml.

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PUM

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Abstract

The invention relates to a chromatographic medium with indole-3-acethydrazide as functional ligand, which includes a chromatographic substrate and the ligand. The chromatographic substrate is hydrophilic porous microspheres having hydroxyl groups, while the ligand is indole-3-acethydrazide which is activated with an epoxy activator and then is coupled. The novel chromatographic medium is designedwith the indole-3-acethydrazide as the functional ligand and has a significant character of hydrophobic charge inducing chromatography. The chromatographic medium has strong adsorption capacity to anantibody and can be used for large-scale production of the antibody.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a chromatographic medium with indole-3-acetylhydrazide as a functional ligand. Background technique [0002] With the rapid development of medicine, some difficult diseases are not as helpless as in the past, such as cancer, rheumatoid arthritis and so on. Surprisingly, the discovery of new targets makes scientists more targeted when designing drugs, greatly improving the success rate of drug screening. At the same time, monoclonal antibodies (referred to as monoclonal antibodies), as biomacromolecule targeting drugs, can be coupled with small molecule drugs to form antibody conjugates ("biomissiles"), which can specifically recognize and bind cancer cells, thereby achieving Efficient therapeutic effect. According to reports, there are 30 types of monoclonal antibodies on the market, which are mainly used for the treatment of cancer and immune system diseases. At present, the numb...

Claims

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Application Information

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IPC IPC(8): B01J20/26B01J20/30C07K1/16
CPCB01J20/26C07K1/16
Inventor 瞿欢欢朱至放周青竹
Owner SUZHOU BOJIN BIOLOGICAL TECH