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Affinity Chromatography Medium Using Tetrapeptide Gastrin as Functional Ligand

A technology of tetrapeptide gastrin and chromatography medium, which is applied in the field of protein chromatography separation technology, can solve the problems of high separation cost and difficult separation cost, and achieves remarkable hydrophobic charge-induced chromatography separation characteristics and adsorption capacity. Large, high ligand density effect

Active Publication Date: 2020-07-14
SUZHOU BOJIN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, 70-80% of the antibody production cost is in the separation process, and the separation cost is difficult to solve through the scale-up of the original technology
The current core technology for antibody separation is Protein A affinity chromatography, which has a good separation effect, but there are also some inherent limitations, resulting in high separation costs and becoming the bottleneck of antibody production.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Get polychitosan 10g, add 25% (v / v) dimethyl sulfoxide of 2g, the sodium hydroxide of 5g divinyl sulfoxide and 3g, activate 10 hours in 200rpm shaking table under 25 ℃, suction filtration , washed with deionized water to obtain an activated chromatography matrix; the activated chromatography matrix was mixed with 10 g of 3-mercaptopropionic acid and 2 g of ammonium persulfate for alcoholization, reacted in a shaking table at 200 rpm at 50 ° C for 3 hours, filtered with suction, and used to remove Wash with ionized water to obtain an alcoholized matrix; mix the alcoholized matrix with 1 g of tetrapeptide gastrin and 0.5M sodium carbonate buffer, the pH of the sodium carbonate buffer is 10, and react in a shaker at 200 rpm at 30°C for 10 hours. The medium after tetrapeptide gastrin coupling was obtained; the medium after tetrapeptide gastrin coupling was suction-filtered, washed with deionized water, added to cyclohexylamine containing 50 g, and reacted in a shaker at 200 ...

Embodiment 2

[0025] Get 10g of polychitosan, add 5g of 25% (v / v) dimethyl sulfoxide, 10g of divinyl sulfoxide and 5g of sodium hydroxide, activate it in a shaker at 200rpm at 25°C for 12 hours, and filter with suction , washed with deionized water to obtain an activated chromatography matrix; the activated chromatography matrix was mixed with 10 g of 3-mercaptopropionic acid and 2 g of ammonium persulfate for alcoholization, reacted in a shaking table at 200 rpm at 50 ° C for 3 hours, filtered with suction, and used to remove Wash with ionized water to obtain alcoholized matrix; mix the alcoholized matrix with 1 g of tetrapeptide gastrin and 0.5M sodium carbonate buffer, the pH of the sodium carbonate buffer is 10, react in a shaker at 200 rpm at 30°C for 8 hours, Obtain the tetrapeptide gastrin-coupled medium; suction filter the tetrapeptide gastrin-coupled medium, wash with deionized water, add to cyclohexylamine containing 10 g, and react in a shaker at 30°C and 200rpm for 7 hours, wash...

Embodiment 3

[0027] Get polychitosan 10g, add 25% (v / v) dimethyl sulfoxide of 2g, the sodium hydroxide of 10g divinyl sulfoxide and the sodium hydroxide of 5g, activate 10 hours in 200rpm shaking table under 25 ℃, suction filtration , washed with deionized water to obtain an activated chromatography matrix; the activated chromatography matrix was mixed with 10 g of 3-mercaptopropionic acid and 3 g of ammonium persulfate for alcoholization, reacted in a shaker at 200 rpm at 50 ° C for 6 hours, filtered with suction, and used to remove Wash with ionized water to obtain an alcoholized matrix; mix the alcoholized matrix with 2g of tetrapeptide gastrin and 0.5M sodium carbonate buffer, the pH of the sodium carbonate buffer is 10, and react in a shaker at 200 rpm at 30°C for 20 hours. The tetrapeptide gastrin-coupled medium was obtained; the tetrapeptide gastrin-coupled medium was suction-filtered, washed with deionized water, added to cyclohexylamine containing 10 g, and reacted in a shaker at 2...

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Abstract

The invention discloses an affinity chromatography medium taking tetragastrin as a functional ligand. The affinity chromatography medium comprises a chromatography substrate and a ligand, wherein thechromatography substrate is poly-chitosan; and the ligand is tetragastrin activated and coupled by diethylene sulfoxide. The novel chromatography medium developed in the invention takes the tetragastrin as the functional ligand, has obvious hydrophobic charge-induced chromatographic separation characteristics, has extremely high adsorption capacity on antibodies and can be applied to large-scale antibody preparation.

Description

technical field [0001] The invention relates to an affinity chromatography medium with tetrapeptide gastrin as a functional ligand, which belongs to the protein chromatography separation technology in the field of biochemical industry. Background technique [0002] Antibodies are hailed as "biological missiles". They have the advantages of strong specificity, good curative effect, and low toxic and side effects. They are mainly used to treat malignant tumors, major infectious diseases, and immune system diseases. It is estimated that the global antibody sales will exceed 50 billion US dollars in 2011, accounting for more than 1 / 3 of the biotechnology drug market. The antibody industry has become the focus of competition for biopharmaceuticals in various countries. However, my country's antibody industry has just started. [0003] The clinical dose of therapeutic antibody is large, the treatment cycle is long, and the high cost of treatment makes ordinary patients daunting. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/24B01J20/30C07K1/22
CPCB01J20/24C07K1/22
Inventor 瞿欢欢朱至放
Owner SUZHOU BOJIN BIOLOGICAL TECH