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Preparation method of high-specific-activity urofollitropin

A technology of urinary follicle-stimulating hormone and high ratio, which is applied in the field of preparation of urinary follicle-stimulating hormone, can solve the problems of increased injection pain and low product purity, and achieve the effects of reduced material cost, improved preparation process conditions, and high purity

Inactive Publication Date: 2013-02-13
RIZHAO XINKANG BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Miscellaneous proteins can cause certain allergic reactions and increase the pain of injection
Therefore, the market tends to develop high-purity FSH with high specific activity recently, but the purity of the products obtained by most of the production processes for preparing FSH is not high.

Method used

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  • Preparation method of high-specific-activity urofollitropin

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preparation example Construction

[0024] A preparation method of high specific activity urinary follicle-stimulating hormone, the specific activity of the high specific activity urinary follicle-stimulating hormone is ≥ 10000 international units / mg protein;

[0025] Described preparation method step is:

[0026] Step 1. Add Tris-HCl equilibrium solution with a pH value of 7.4 to 20-60 g of low-purity urotropin to prepare a 5-15 L mixed solution, and then pass the mixed solution through 5 L of anti-luteinizing hormone antibody gel Column, collect the adsorbed liquid to obtain the adsorbed liquid, and then use 20-60L Tris-HCl equilibrium solution with a pH value of 7.4 to elute, collect the eluent I, and finally combine the eluent I and the adsorbed liquid to obtain a mixed liquid, and then the mixed solution was purified and concentrated through an ultrafiltration membrane with a molecular weight cut-off of 10KD to obtain a concentrated solution of 5-15 L for subsequent use;

[0027] Step 2. Adjust the pH valu...

Embodiment 1

[0035] A preparation method of high specific activity urinary follicle-stimulating hormone, the preparation method steps are:

[0036] Step 1. Add Tris-HCl balance solution with a pH value of 7.4 to 20 g of low-purity urotropin, wherein the biological potency of follicle-stimulating hormone is 60 IU / mg, and the biological potency of luteinizing hormone is 55 IU / mg. Mix evenly to prepare 5L of mixed liquid, and then pass the mixed liquid through a 5L anti-luteinizing hormone antibody gel column, luteinizing hormone is combined with the antibody on the anti-luteinizing hormone antibody gel column, and luteinizing hormone is adsorbed on the anti-luteinizing hormone antibody gel column. The follicle-stimulating hormone remained on the adsorbed liquid, and the adsorbed liquid was collected, and then eluted with 20L of Tris-HCl equilibrium solution with a pH value of 7.4, and the eluate I was collected, and then Combine the eluent I and the adsorption solution to obtain a mixed solu...

Embodiment 2

[0042] A preparation method of high specific activity urinary follicle-stimulating hormone, the preparation method steps are:

[0043] Step 1. Add Tris-HCl balance solution with a pH value of 7.4 to 40 g of low-purity urotropin, wherein the biological potency of follicle-stimulating hormone is 65 IU / mg, and the biological potency of luteinizing hormone is 60 IU / mg. mg, mixed evenly to obtain 10L of mixed liquid, and then the mixed liquid was passed through a 10L anti-luteinizing hormone antibody gel column, luteinizing hormone was combined with the antibody on the anti-luteinizing hormone antibody gel column, and luteinizing hormone was adsorbed on Anti-luteinizing hormone antibody gel column, while follicle-stimulating hormone remains in the adsorbed liquid, collect the adsorbed liquid, and then use 40L of Tris-HCl equilibrium solution with a pH value of 7.4 to elute, and collect the eluent Ⅰ , and finally the eluent I and the adsorption solution were mixed to obtain a mixed ...

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Abstract

The invention relates to a preparation method of high-specific-activity urofollitropin, which comprises the following steps: adding a Tris-HCl equilibrium liquid with the pH value of 7.4 into low-purity menotrophin, passing through an anti-luteinizing-hormone antibody gel column, purifying and concentrating; sequentially carrying out anion-exchange chromatography, cation-exchange chromatography and blue sepharose dye affinity chromatography; collecting the washing liquid, and detecting washing liquid with a 280nm ultraviolet spectrophotometer; and when the OD (optical density) value is 0.002, collecting the washing liquid, purifying, concentrating, precipitating with absolute ethanol, and dehydrating to obtain the high-specific-activity urofollitropin of which the specific activity is higher than or equal to 10000 international units / mg protein. The preparation technique provided by the invention is simple and convenient to operate; the specific activity of the prepared urofollitropin is higher than or equal to 10000 international units / mg protein; and the product has high activity and high purity.

Description

technical field [0001] The invention relates to a preparation method of urinary follicle-stimulating hormone, in particular to a preparation method of high specific activity urinary follicle-stimulating hormone. Background technique [0002] Follicle-stimulating hormone (FSH) is a type of glycoprotein synthesized and secreted by basophils in the anterior pituitary gland. It consists of two subunits, the α chain and the β chain. The α subunit has 85 amino acids, and the β subunit has 115 amino acids. The specificity is composed of β subunits, and the total molecular weight is about 33,000. The β subunit of FSH consists of 115 amino acids with a molecular weight of about 18,000, and the asparagine at the 7th and 24th positions are amino acids that undergo N-glycosylation. [0003] The main physiological function of follicle-stimulating hormone (FSH) is to promote the growth and development of various cells of the follicle: to promote the differentiation of mesenchymal cells ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/59C07K1/36C07K1/34C07K1/22C07K1/18
Inventor 申恒海钱兴泽梁云爱杜安生
Owner RIZHAO XINKANG BIOLOGICAL TECH
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