Unlock instant, AI-driven research and patent intelligence for your innovation.

A kind of preparation method of high-purity urinary follicle-stimulating hormone

A high-purity technique for urofollitropin, which is applied in the field of extraction and preparation of urofollitropin, can solve the problems of high price of antibody immunoaffinity chromatography column, contamination of heterologous proteins, hindering application and promotion, etc.

Active Publication Date: 2020-01-03
江西浩然生物制药有限公司
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, most antibodies are of mouse or rabbit origin, which are proteins themselves. During the chromatography process, the shedding of antibodies coupled to the matrix will cause contamination of heterologous proteins and affect product quality.
At the same time, the antibody immunoaffinity chromatography column is expensive, has a short service life, and is difficult to maintain and industrially scale up production, all of which seriously hinder its application and promotion.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of preparation method of high-purity urinary follicle-stimulating hormone
  • A kind of preparation method of high-purity urinary follicle-stimulating hormone
  • A kind of preparation method of high-purity urinary follicle-stimulating hormone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Such as figure 1 As shown, the crude product of HMG (manufactured by our company, batch number Z0203150901) is used as the starting material, its FSH biological potency is 30IU / mg, LH biological potency is 28IU / mg, and FSH specific activity is 43IU / mg protein.

[0040] 1. Capto DEAE anion exchange chromatography

[0041] Dissolve 8.0g HMG crude product with 200ml water, adjust the pH value of solution to 7.5, this solution loading sample is to the Capto DEAE chromatographic column (purchased from GE company) that column volume is 250ml, chromatographic column is pre-used with equilibrium liquid (0.02 M PB, pH7.5) is well balanced; after loading the sample, wash the unadsorbed protein with the equilibrium solution, and detect the absorbance value of the washing solution at 280nm until the absorbance value no longer decreases; then use the eluent (0.02M PB +0.15MNaCl) to elute FSH, and detect the absorbance value of the eluent at 280nm until the absorbance value no longe...

Embodiment 2

[0052] Using low-purity HMG (manufactured by our company, batch number Z0203150901) as the starting material, its FSH biological potency is 30IU / mg, LH biological potency is 28IU / mg, and FSH specific activity is 43IU / mg protein.

[0053] 1. Capto DEAE anion exchange chromatography

[0054] Dissolve 20g HMG with 500ml of water, adjust the pH value of solution 1 to 7.5, this solution loads the sample to the Capto DEAE chromatographic column (purchased from GE company) that column volume is 630ml, chromatographic column is pre-used with equilibrium liquid (0.02M PB , pH7.5) well balanced; after loading the sample, wash the unadsorbed protein with the balance solution, and detect the absorbance value of the washing solution at 280nm until the absorbance value no longer decreases; then use the eluent (0.02M PB+0.15 M NaCl) to elute FSH, and detect the absorbance value of the eluate at 280nm until the absorbance value no longer decreases, collect a total of 2.5L of the eluate efflue...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a preparation method of high-purity urine follicle stimulating hormone (u-FSH). The preparation method comprises the following steps: (1) carrying out anion exchange chromatography purification on a human menopausal gonadotropin (HMG) crude product to obtain a urine follicle stimulating hormone intermediate I; (2) carrying out cathion exchange chromatography purification on the urine follicle stimulating hormone intermediate I to obtain a urine follicle stimulating hormone intermediate II; (3) carrying out dye affinity chromatography purification on the urine follicle stimulating hormone intermediate II to obtain a urine follicle stimulating hormone intermediate III; (4) carrying out hydrophobic chromatography purification on the urine follicle stimulating hormone intermediate III to obtain the high-purity urine follicle stimulating hormone. The method provided by the invention has the advantages of simplicity and convenience in operation, low production cost, higher purity of follicle stimulating hormone (FSH), lower content of luteotropic hormone (LH) and no heterologous protein pollution.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for extracting and preparing urinary follicle-stimulating hormone. Background technique [0002] Urine Follicle Stimulating Hormone (u-FSH) is a composition whose main component is follicle stimulating hormone (Follicle Stimulating Hormone, FSH). The FSH content in high-purity urinary follicle stimulating hormone can reach more than 95%, and it hardly contains luteinizing hormone ( Luteotropic Hormone, LH). FSH is a glycoprotein hormone with a molecular weight of about 30KD. FSH can promote the proliferation of granulosa cells, stimulate steroid production, regulate the development and maturation of gametocytes, and is one of the main hormones in the hypothalamus-pituitary-gonad axis. FSH has the functions of regulating menstrual cycle, ovulation and regulating male testicular function in humans. It is commonly used clinically to treat female infertility, insufficient secr...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/59C07K1/36C07K1/34C07K1/22C07K1/20C07K1/18
CPCC07K14/59
Inventor 万偲魏超娟徐富勇黄优生胡著阳李菊根
Owner 江西浩然生物制药有限公司