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Chorionic gonadotrophin purification method

A technology for chorionic gonadotropin and purification method, which is applied in the field of chorionic gonadotropin purification, can solve the problems of low yield, complicated process, unsatisfactory potency, etc., and achieves high purity, simplified purification process, and reduced floor space. Effect

Pending Publication Date: 2016-09-28
宁波人健药业集团股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the process is complicated, the yield is low, and the highest potency is only about 5000iu / mg
Such as CN104497128A, provides a kind of low-salt extraction method to extract chorionic gonadotropin method from crude product, but the potency of this method is also unsatisfactory

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] A method for purifying chorionic gonadotropin, comprising the following steps:

[0097] (1) The crude chorionic gonadotropin is dissolved in 1% sodium acetate solution, after stirring, add 100% alcohol to adjust the alcohol concentration to 50%, after stirring and sedimenting, take the supernatant, add 100% alcohol to the supernatant to adjust the alcohol concentration to 85 %, take the precipitate, centrifuge, and dry under reduced pressure at room temperature to obtain a solid with a potency of about 200iu / mg.

[0098] (2) The first step of chromatography: the solid is dissolved in equilibrium solution 1, and the protein solution A is initially obtained by GE capto blue (high sub) affinity chromatography.

[0099] Equilibrium solution 1: 50 nmol tris+ 100 nmol NaCl, pH 7.5;

[0100] Wash solution 1: 50 nmol tris + 500 nmol NaCl + 100% ethanol to make the final concentration 5%, pH 7.5;

[0101]Eluent 1: 50 nmol tris+ 2 mol NaCl+100% ethanol to make the final concent...

Embodiment 2

[0127] A method for purifying chorionic gonadotropin, comprising the following steps:

[0128] (1) Crude chorionic gonadotropin was dissolved in 1% sodium acetate solution, and after stirring, 100% alcohol was added to adjust the alcohol concentration to 50%. %, take the precipitate, centrifuge, and dry under reduced pressure at room temperature to obtain a solid with a potency of about 200iu / mg.

[0129] (2) The first step of chromatography: the solid is dissolved in equilibrium solution 1, and the protein solution is initially obtained by GE capto blue (high sub) affinity chromatography.

[0130] Equilibrium solution 1: 50 nmol tris+ 100 nmol NaCl, pH 7.5;

[0131] Wash solution 1: 50 nmol tris+ 500 nmol NaCl+100% ethanol to make the final concentration 5%, pH 7.5;

[0132] Eluent 1: 50 nmol tris+ 2 mol NaCl+100% ethanol to make the final concentration 20%, pH 7.5;

[0133] Equilibrium solution 1 equilibrates, the sample is loaded on the column, balance solution 1 rebalan...

Embodiment 3

[0150] A method for purifying chorionic gonadotropin, comprising the following steps:

[0151] (1) Crude chorionic gonadotropin was dissolved in 1% sodium acetate solution, and after stirring, 100% alcohol was added to adjust the alcohol concentration to 50%. %, take the precipitate, centrifuge, and dry under reduced pressure at room temperature to obtain a solid with a potency of about 200iu / mg.

[0152] (2) The first step of chromatography: the solid is dissolved in equilibrium solution 1, and the protein solution is initially obtained by GE capto blue (high sub) affinity chromatography.

[0153] Equilibrium solution 1: 50 nmol tris+ 100 nmol NaCl, pH 7.5;

[0154] Wash solution 1: 50 nmol tris + 500 nmol NaCl + 100% ethanol to make the final concentration 5%, pH 7.5;

[0155] Eluent 1: 50 nmol tris+ 2 mol NaCl+100% ethanol to make the final concentration 20%, pH 7.5;

[0156] Equilibrium solution 1 equilibrates, the sample is loaded on the column, balance solution 1 reba...

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PUM

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Abstract

The invention relates to a chorionic gonadotrophin purification method which adopts three times of chromatography for purification, wherein the three times of chromatography includes affinity chromatography, hydrophobic chromatography and ion-exchange chromatography, and finally, a high-purity chorionic gonadotrophin solid is obtained. The filler of the affinity chromatography is capto blue (high sub), the filler of the hydrophobic chromatography is Phenyl Sepharose 6 Fast Flow (high sub), and the filler of the ion-exchange chromatography is DEAE Sepharose Fast Flow. According to the chorionic gonadotrophin purification method, the purification process is optimized, the purification flows are simplified, the yield is obviously increased, the titer is higher than that of the existing process, the floor area of the equipment is reduced, and the energy consumption of a workshop is reduced. In the invention, the titer of preprocessed chorionic gonadotrophin is about 200iu / mg, and the titer of chorionic gonadotrophin processed by three times of chromatography can reach 10000iu / mg, so that the titer is obviously increased.

Description

technical field [0001] The invention belongs to the technical field of medicine and relates to a method for purifying chorionic gonadotropin. Background technique [0002] Chorionic Gonadotrophin is a natural gonadotropin extracted and refined from the urine of healthy pregnant women, which has a function similar to that of luteinizing hormone. For women, it can promote the maturation of follicles and ovulation, and maintain the function of the corpus luteum. For men, it can stimulate the development of Leydig cells in the testis, increase the secretion of androgen, promote the descent of the testis and the development of male secondary sexual characteristics. Combined with urogonadotropin, it can simulate the physiological peak of luteinizing hormone and trigger ovulation. Clinically, chorionic gonadotropin is still the drug of choice for the treatment of menstrual disorders, anovulatory infertility, threatened abortion and habitual abortion, cryptorchidism, and male sexu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/59C07K1/22C07K1/20C07K1/18
CPCC07K14/59
Inventor 夏星辉胡晶锦王群芳
Owner 宁波人健药业集团股份有限公司
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