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Method for extracting gonadotrophic hormone from human urine
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A gonadotropin and urine technology, which is applied in the field of extracting gonadotropin from human urine, can solve the problems of low purity, low product yield, and high impurity content in the product, and achieve simple process technology, good product purity, and high product quality. The effect of high yield
Inactive Publication Date: 2015-10-07
厦门欧瑞捷生物科技有限公司
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Problems solved by technology
[0005] At present, the extraction process of urinary follicle-stimulating hormone and chorionic gonadotropin is relatively complicated, the production cycle is long, and the product yield is not high, which leads to a great increase in production cost and cannot meet market demand
Yin Xiaowei etc. disclosed "a kind of extraction method of urinary follicle-stimulating hormone" in the Chinese invention patent whose publication number is CN101555278A; The method of "; the extraction process disclosed in these two patent applications, compared with the prior art, has the advantages of simple process technology and high product yield, but also has the disadvantages of high impurity content, low purity, and low potency. Resulting in cumbersome subsequent refining process steps
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Embodiment 1
[0019] Embodiment 1, the extraction of urotropin, comprises the following steps:
[0020] (1) Collect 1000L of urine from menopausal women, put the urine that passes the test into the tank, and adjust the pH value of the urine to 8.0-9.0 with alkali;
[0021] (2) After adjusting the pH value, add 10kg of diatomaceous earth to the urine, stir evenly, and filter with a plate frame;
[0022] (3) Pour the filtered solution into the ultrafiltration tank, filter through the ultrafiltration membrane with a molecular pore size of 10,000, and pour 10L of the concentrated solution into the precipitation tank;
[0023] (4) After prepacking the CM SepharoseFast Flow filler into the ion exchange chromatography column, equilibrate with 2 times the column volume of pH 4.5 NaAc buffer A, put the concentrated solution on the column, and then wash it with 2 times the column volume of buffer A, Then elute with 2 times column volume of buffer B containing 0.3M NaCl, and collect the eluate;
[...
Embodiment 2
[0027] Embodiment 2, the extraction of chorionic gonadotropin, comprises the following steps:
[0028] (1) Use 1000L of urine raw material from pregnant women, pump the urine that has passed the inspection into the tank, and adjust the pH value of the urine to 8.0-9.0 with alkali;
[0029] (2) After adjusting the pH value, add 10kg of diatomaceous earth or kaolin to the urine, stir for 30 minutes until uniform, and filter with a plate frame;
[0030] (3) Pump the filtered solution into the ultrafiltration tank, filter through the ultrafiltration membrane with a molecular weight cut-off of 10,000, and put 10L of the obtained ultrafiltration concentrate into the precipitation tank;
[0031] (4) After prepacking the SepharoseFast Flow filler into the ion exchange chromatography column, equilibrate with 2 times the column volume of pH 4.5 NaAc buffer A, put the concentrated solution on the column, and then wash it with 2 times the column volume of buffer A, and then Elute with 2...
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Abstract
The invention discloses a method for extracting gonadotrophic hormone from human urine. The method is characterized by comprising the following steps: step 1, adjusting a pH value so as to allow impurities to be precipitated; step 2, carrying out filtering to remove the impurities and then carrying out ultrafiltration and concentration; step 3, carrying out ion exchange column chromatography; step 4, carrying out precipitation and impurity removal; and step 5, carrying out precipitation and drying so as to obtain a crude gonadotrophic hormone product. The method for extracting gonadotrophic hormone from human urine has the advantages of simple process, high product yield, low production cost, good product purity, high titer, etc. and enables a plurality of steps to be omitted in subsequent extraction of a high purity product.
Description
technical field [0001] The invention relates to a process for extracting chemical drugs, in particular to a method for extracting gonadotropin from human urine. Background technique [0002] Gonadotropins are glycoprotein hormones that regulate the development of vertebrate gonads and promote the production and secretion of sex hormones. For example, urinary gonadotropin (HMG) secreted by the anterior pituitary gland, human chorionic gonadotropin (HCG, referred to as chorionic gonadotropin) secreted by human placenta, and luteinizing hormone (LH) are all widely used in infertility. They all have a similar structure, consisting of two peptide chains, α and β, and the sequences of the α chains are completely identical. The difference between different proteins is in the sequence of β chain, which is also reflected in different spatial structure and binding to different cell surface receptors. [0003] Urogonadotropin (HMG) is a type of reproductive hormone extracted from th...
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