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Purifying and producing process for high purity follicle stimulating hormone in urine

A follicle-stimulating hormone and purification method technology, which is applied in the field of purification of high-purity urinary follicle-stimulating hormone, can solve the problems of FSH variability, easy loss, and unsuitability for industrial production, and achieve the effect of reducing costs

Inactive Publication Date: 2008-09-10
NANCHANG WANHUA BIOCHEM PHARMA
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Problems solved by technology

In this patent, due to the use of antibodies for affinity chromatography, and the affinity of the antibodies to their corresponding antigen FSH is very high, the elution must use strong alkaline conditions (ammonia) to elute the FSH bound to the agarose. Under these conditions, FSH is easily denatured into a protein without biological activity, or easily loses its sugar groups and becomes a substance that strongly inhibits normal physiological functions. Therefore, the process involved in this patent is not suitable for industrial production

Method used

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  • Purifying and producing process for high purity follicle stimulating hormone in urine
  • Purifying and producing process for high purity follicle stimulating hormone in urine

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Embodiment Construction

[0011] Coupling of red dye and Sepharose 4B: Weigh a certain amount of red dye (azo dyes, such as Reactive Red 120) ligand and add it to distilled water, so that the initial ligand concentration is 0.5~3.5% (W / V) In between, stir to dissolve, then add 10-50% (W / V) to wash and filter the dried Sepharose 4B gel, place it on a constant temperature water bath shaker and raise it to the desired reaction temperature (40-80℃) within 1-10min , Then add dye auxiliaries (such as NaCl), and then add Na after 30 minutes 2 CO 3 (The final concentration is 10-25g / L) Fix for 15min, then add catalyst (such as DABCO) and react for 2-10 hours. The pH of the reaction is controlled between 9-10. The coupled gel is repeatedly washed with pure water for use.

[0012] FSH purification step 1-crude product processing: the FSH activity in the crude HMG product is about 1IU / mg, and the activity of LU is about 1IU / mg. Since the crude HMG contains a lot of impurities, as well as impurities such as kaolin, de...

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Abstract

During the purification and production of high purity uric follicle stimulating hormone, red azo dye is coupled to Sepharose 4B as the medium of affinity separating and purifying uric follicle stimulating hormone, and through dye affinity chromatography, QAE ion exchange chromatography and other technological steps, high purity uric follicle stimulating hormone product is obtained from uric gonadotropin from post-menopause. The product produced based on the said technological process has biological potency not lower than 200 IU / mg and FSH / LH higher than 100, is white and soluble, and may be used directly in producing injection.

Description

Technical field [0001] The invention relates to a purification method of high-purity urinary follicle stimulating hormone, in particular to a purification method of obtaining a follicle stimulating hormone (FSH) product from crude human menopausal gonadotropin (HMG). Background technique [0002] Follicle Stimulating Hormone (FSH), also known as Follicle Stimulating Hormone, is a glycoprotein synthesized and secreted by pituitary basophils. It has a molecular weight of 33,000 Dal and consists of two non-covalent α and β subunits. The α subunit contains 92 amino acids. It is compatible with Luteinizing hormone (LH) and Human Chorionicgonadotropin (HCG) The subunits are very similar. The β subunit is composed of 115 amino acids and is the component that determines its specificity. Because FSH is a glycoprotein hormone, and its molecular weight, isoelectric point and other physical and chemical properties are very similar to LH, TSH (thyroid stimulating hormone), etc., classical ion...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/59C07K1/18
Inventor 张万山刘兰花李菊根王启要
Owner NANCHANG WANHUA BIOCHEM PHARMA
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