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Method for improving chorionic gonadotrophin titer

A technology of chorionic gonadotropin and purification method, which is applied in the direction of chemical instruments and methods, decapeptides, peptides, etc., can solve the problems of low potency of chorionic gonadotropin products and cumbersome process flow, etc., and achieve the reduction of floor area and optimization The purification process and the effect of reducing energy consumption in the workshop

Inactive Publication Date: 2017-05-10
青岛冠龙生物制药有限公司
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chorionic gonadotropin is the main export biochemical drug in my country. There are many methods for purifying chorionic gonadotropin, but the current method has a complicated process and low potency of chorionic gonadotropin products.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] (1) Pretreatment: Dissolve the crude chorionic gonadotropin in the dissolving solution, then add ethanol solution until there is a low concentration of ethanol in the mixed solution, then precipitate, take the supernatant, add the ethanol solution to the mixed solution In the presence of high concentration of ethanol, take the precipitated solid;

[0017] (2) The first step of chromatography: the solid obtained in step (1) is subjected to capto blue (high sub) affinity chromatography to obtain protein solution A;

[0018] (3) The second step of chromatography: the protein solution A obtained in step (2) is subjected to Phenyl Sepharose 6 Fast Flow (highsub) hydrophobic chromatography to obtain protein solution B;

[0019] (4) The third step of chromatography: the protein solution B obtained in step (3) was subjected to DEAE Sepharose Fast Flow ion exchange chromatography to obtain protein solution C;

[0020] (5) Post-treatment: add ethanol solution to the protein solu...

Embodiment 2

[0022] (1) Pretreatment: Dissolve the crude chorionic gonadotropin in the dissolving solution, then add ethanol solution until there is a low concentration of ethanol in the mixed solution, then precipitate, take the supernatant, add the ethanol solution to the mixed solution In the presence of high concentration of ethanol, take the precipitated solid;

[0023] (2) The first step of chromatography: the solid obtained in step (1) is subjected to capto blue (high sub) affinity chromatography to obtain protein solution A;

[0024] (3) The second step of chromatography: the protein solution A obtained in step (2) is subjected to Phenyl Sepharose 6 Fast Flow (highsub) hydrophobic chromatography to obtain protein solution B;

[0025] (4) Post-treatment: add ethanol solution to the protein solution C obtained in step (3) until there is a high concentration of ethanol in the mixed solution, and collect the solid after precipitation to obtain chorionic gonadotropin solid.

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Abstract

The invention discloses a method for purifying chorionic gonadotrophin. The technology is capable of using three times of chromatography to purify the chorionic gonadotrophin of low titer, and purifying the chorionic gonadotrophin from about 300 IU / mg of the titer to about 9000 IU / mg of the titer. The titer is greatly improved and other quality standards accord with each index specified by Chinese Pharmacopoeia, United States Pharmacopeia and European Pharmacopoeia. The method has the advantages of simpliness, high product purity and yield, and low cost, and is suitable for large-scale industrial production of the pharmaceuticals industry.

Description

technical field [0001] The invention belongs to the field of pharmaceutical production, and specifically illustrates a method for purifying chorionic gonadotropin by using three chromatography techniques. Background technique [0002] Chorionic gonadotropin is a biologically active substance extracted from the urine of pregnant women, which has the functions of promoting embryonic development and maintaining pregnancy. Chorionic gonadotropin, thyroid-stimulating hormone, follicle-stimulating hormone, and luteinizing hormone are members of the glycoprotein hormone family. Clinically, chorionic gonadotropin can treat diseases such as habitual abortion and infertility caused by excessive secretion of follicle-stimulating hormone or insufficient secretion of luteinizing hormone. In addition, it is also widely used in aquaculture and animal husbandry production. Chorionic gonadotropin is the main export biochemical drug in my country. There are many methods for purifying chorio...

Claims

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Application Information

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IPC IPC(8): C07K14/59C07K1/22C07K1/20C07K1/18
CPCC07K14/59
Inventor 刘乃山马雪陈娥功
Owner 青岛冠龙生物制药有限公司
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