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A kind of purification method of urinary gonadotropin

A gonadotropin and purification method technology, which is applied in the field of purification of urinary gonadotropin, can solve the problems of inability to effectively protect the biological activity of the product, low immunity and biological potency, and large loss of activity, etc., and the process conditions are easy to control , high biological potency and low cost

Active Publication Date: 2019-07-02
SHANDONG TOPSCI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In actual operation, the activity recovery rate of the product obtained by this extraction method is between 70% and 80%, and the biggest defect is that the total activity loss is large, the immunity and biological titer of the product are low, and the biological activity of the product cannot be effectively protected.

Method used

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  • A kind of purification method of urinary gonadotropin

Examples

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Embodiment 1

[0027] (1) Preparation of polyethylene glycol ether-ammonium sulfate two-phase aqueous system: add ammonium sulfate to water at 15°C to prepare an ammonium sulfate solution, then add polyethylene glycol ether with a molecular weight of 2000 to the ammonium sulfate solution, and mix well , to obtain a polyethylene glycol ether-ammonium sulfate two-phase system solution, wherein the mass percent of polyethylene glycol ether in the two-phase system 1 is 10%, and the mass percent of ammonium sulfate is 10%; in the two-phase system 2 Polyethylene glycol ether mass percentage is 10%, and the mass percentage of ammonium sulfate is 18%;

[0028] (2) Pass 100,000L of the raw material solution through an ultrafiltration membrane with a molecular weight cut-off of 10,000 to obtain an ultrafiltration concentrate. Keep the temperature of the ultrafiltration concentrate at 15°C, add an equal volume of two-phase aqueous system 1 and stir evenly, and centrifuge at 1000r / min for 5min , to obta...

Embodiment 2

[0035](1) Preparation of polyethylene glycol ether-ammonium sulfate two-phase system: add ammonium sulfate to water at 20°C to prepare ammonium sulfate solution, then add polyethylene glycol ether with a molecular weight of 2000 to the solution, mix well, and prepare Obtain polyethylene glycol ether-ammonium sulfate two-phase system solution, wherein the mass percent of polyethylene glycol ether in two-phase system 1 is 20%, and the mass percent of ammonium sulfate is 15%; Glycol ether mass percentage is 20%, and the mass percentage of ammonium sulfate is 24%;

[0036] (2) 100,000L of raw material solution is passed through an ultrafiltration membrane with a molecular weight cut-off of 10,000 to obtain an ultrafiltration concentrate. Keep the temperature of the ultrafiltration concentrate at 20°C, add an equal volume of two-phase aqueous system 1 and stir evenly, and centrifuge at 2000r / min for 2min , to obtain a polyethylene glycol ether organic phase containing urinary gonad...

Embodiment 3

[0043] (1) Preparation of polyethylene glycol ether-ammonium sulfate two-phase system: add ammonium sulfate to water at 20°C to prepare ammonium sulfate solution, then add polyethylene glycol ether with a molecular weight of 2000 to the solution, mix well, and prepare Obtain polyethylene glycol ether-ammonium sulfate two-phase system solution, wherein the mass percent of polyethylene glycol ether in two-phase system 1 is 14%, and the mass percent of ammonium sulfate is 13%; Glycol ether mass percentage is 17%, and the mass percentage of ammonium sulfate is 20%;

[0044] (2) 100,000L of raw material solution is passed through an ultrafiltration membrane with a molecular weight cut-off of 10,000 to obtain an ultrafiltration concentrate. Keep the temperature of the ultrafiltration concentrate at 20°C, add an equal volume of two-phase aqueous system 1 and stir evenly, and centrifuge at 2000r / min for 2min , to obtain a polyethylene glycol ether organic phase containing urinary gona...

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Abstract

The invention relates to the field of pharmacy, in particular to a purification method for urinary gonadotropin.After urine of a postmenopausal woman is taken and subjected to ultrafiltration and concentration and ultrafiltrate is extracted two times with a polyglycol ether-ammonium sulfate aqueous two-phase system, an organic phase containing urinary gonadotropin is obtained, the organic phase is treated in a water bath, split and centrifuged, an aqueous phase containing urinary gonadotropin is obtained, the aqueous phase passes through an ultrafiltration membrane and then is fed to a hydroxyapatite chromatographic column, eluant is collected, an ethanol solution is added to the eluant, the alcoholic strength is adjusted, standing and filtering are conducted, sediment is obtained, and the purified urinary gonadotropin is obtained after drying.According to the purification method for the urinary gonadotropin, the biological value of follicle-stimulating hormones and luteinizing hormones in the obtained urinary gonadotropin is high, the activity recovery rate is high, the method is easy to operate, and industrial production is easy.

Description

technical field [0001] The invention relates to the field of pharmacy, in particular to a method for purifying urinary gonadotropin. Background technique [0002] Urinary gonadotropin (HMG) contains two active ingredients, follicle stimulating hormone (FSH) and luteinizing hormone (LH), which are glycoprotein hormones produced by the pituitary gland, and are composed of two subunits of α-chain and β-chain . Clinically, urinary gonadotropin is mainly used to treat infertility. It can be extracted from the pituitary gland or the urine of menopausal women, and can also be prepared by DNA recombinant technology. [0003] Because the level of endogenous urinary luteinizing hormone is different in different patients, it is clinically necessary to use preparations with different ratios of urinary follicle-stimulating hormone and urinary luteinizing hormone to suit the case. The clinical application of urinary gonadotropin is mainly realized by chromatographic separation technolog...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/59C07K1/34
CPCC07K14/59
Inventor 潘涛张富源邢仕杰殷晓薇
Owner SHANDONG TOPSCI BIO TECH