Purification method based on gonadotropin in urine

A gonadotropin and purification method technology, which is applied in the field of gonadotropin purification based on urine, can solve the problems of rash, pain of gonadotropin injection preparations, different gonadotropin components, etc., and achieves improved purity and high titer. , the effect of effective separation

Pending Publication Date: 2022-07-29
JIANGSU YOULIKA BIOTECHNOLOYG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the gonadotropins extracted from human urine in CN105732797B or other prior art have more heterogeneous chemical components, and the components of gonadotropins e...

Method used

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  • Purification method based on gonadotropin in urine
  • Purification method based on gonadotropin in urine
  • Purification method based on gonadotropin in urine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] A method for purifying gonadotropins in urine, comprising the following steps:

[0028] S1. Collect urine for dialysis

[0029] Urine dialysis of menopausal women with purified water for 3h, 13mM Tris-HCl buffer solution with pH value of 7.9 for dialysis of urine for 3.5h, centrifugation to obtain supernatant;

[0030] In the present embodiment, the volume ratio of the purified water, Tris-HCl buffer solution and urine is 2:2:1;

[0031] S2. Chromatography to collect the eluate

[0032] The supernatant obtained in step S1 was placed on a hydroxyapatite chromatographic column, and the 0.05M acetic acid-sodium acetate solution with a pH of 5.6 was used to wash 4 column volumes, and then the 0.1M containing 0.2M sodium chloride with a pH of 5.6 was used. Acetic acid-sodium acetate elution, detect the absorbance value of the eluate at 280nm, and collect the eluate whose absorbance value is greater than 0.1;

[0033] S3. Separation

[0034] To the eluent in step S2, slow...

Embodiment 2

[0040] A method for purifying gonadotropins in urine, comprising the following steps:

[0041] S1. Collect urine for dialysis

[0042] Urine dialysis of menopausal women with purified water for 4h, 13mM Tris-HCl buffer solution with pH value of 7.7 for dialysis of urine for 3h, centrifugation to obtain supernatant;

[0043] In the present embodiment, the volume ratio of the purified water, Tris-HCl buffer solution and urine is 1:3:1;

[0044] S2. Chromatography to collect the eluate

[0045] The supernatant obtained in step S1 was placed on a hydroxyapatite chromatographic column, and the 0.05M acetic acid-sodium acetate solution with a pH of 6.0 was used to wash 3 column volumes, and then the 0.1M containing 0.2M sodium chloride with a pH of 6.0 was passed through. Acetic acid-sodium acetate elution, detect the absorbance value of the eluate at 280nm, and collect the eluate whose absorbance value is greater than 0.1;

[0046] S3. Separation

[0047] To the eluent in step ...

Embodiment 3

[0053] A purification method based on gonadotropins in urine, comprising the following steps:

[0054] S1. Collect urine for dialysis

[0055] Urine dialysis of menopausal women with purified water for 4 hours, 14mM Tris-HCl buffer solution with pH value of 8.1 for 4 hours of urine dialysis, centrifugation to obtain supernatant;

[0056] In the present embodiment, the volume ratio of the purified water, Tris-HCl buffer solution and urine is 3:1:1;

[0057] S2. Chromatography to collect the eluate

[0058] The supernatant obtained in step S1 was placed on a hydroxyapatite chromatographic column, and the 0.05M acetic acid-sodium acetate solution with a pH of 5.5 was used to wash 5 column volumes, and then the 0.1M containing 0.2M sodium chloride with a pH of 5.5 was passed through. Acetic acid-sodium acetate elution, detect the absorbance value of the eluate at 280nm, and collect the eluate whose absorbance value is greater than 0.1;

[0059] S3. Separation

[0060] To the elu...

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Abstract

The invention relates to the technical field of pharmacy, in particular to a purification method based on gonadotropin in urine. The invention relates to a purification method based on gonadotropin in urine. The purification method comprises the following steps: S1, collecting urine and carrying out dialysis treatment; s2, carrying out chromatography and collecting an eluent; s3, separating; and S4, further processing. According to the purification method based on the gonadotropin in the urine, the gonadotropin obtained through purification has the advantages of being high in titer and purity, and the prepared gonadotropin injection preparation does not cause adverse reactions such as pain or erythema.

Description

technical field [0001] The invention relates to the field of pharmaceutical technology, in particular to a method for purifying gonadotropins in urine. Background technique [0002] Gonadotropins are glycoprotein hormones that regulate the development of vertebrate gonads and promote the production and secretion of sex hormones, such as follicle-stimulating hormone (FSH) and luteinizing hormone (LH) secreted by the anterior pituitary gland. The development of germ cells in the testis and the production and secretion of sex hormones; the human placenta secretes chorionic gonadotropin (HCG), which can promote the secretion of progesterone from the corpus luteum of pregnancy. HCG can appear in the urine in the early stage of pregnancy, and it peaks at two months of pregnancy. It is often used as a pregnancy indicator in clinical practice. [0003] Due to the lack of estrogen and progesterone in postmenopausal women, the pituitary gland is feedback-regulated to secrete a large ...

Claims

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Application Information

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IPC IPC(8): C07K14/59C07K1/14C07K1/16C07K1/34C07K1/36
CPCC07K14/59
Inventor 顾京唐维熊心磊王德军李争徐来红
Owner JIANGSU YOULIKA BIOTECHNOLOYG CO LTD
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