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

A follicle-stimulating hormone and production process technology, which is applied in the field of follicle-stimulating hormone products through purification, can solve the problems of FSH variability, no biological activity, and unsuitability for industrial production, and achieve the effect of reducing costs

Inactive Publication Date: 2005-03-02
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

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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 dye, 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 Sepharose 4B gel, place it in a constant temperature water bath shaker and rise to the required reaction temperature (40-80°C) within 1-10min , then add dyeing aids for dyeing (such as NaCl), and then add NaCl after 30 minutes 2 CO 3 (The final concentration is 10-25g / L) fix for 15min, then add a catalyst (such as DABCO) and react for 2-10 hours. The pH of the reaction was controlled between 9-10. The coupled gel was washed repeatedly with pure water for later use.

[0012] Purification step 1 of FSH - treatment of crude product: the activity of FSH in crude HMG is about 1 IU / mg, and the activity of LU is about 1 IU / mg. Because the HMG crude product contains a lot of miscellaneous proteins, ...

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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 and production process of high-purity urinary follicle-stimulating hormone, in particular to a process for obtaining follicle-stimulating hormone (FSH) product by purifying urinary gonadotropin (HMG) crude product. Background technique [0002] Follicle Stimulating Hormone (FSH), also known as follicle-stimulating hormone, is a glycoprotein synthesized and secreted by pituitary basophils. Its molecular weight is 33,000Dal, composed of two non-covalent α and β subunits, the α subunit contains 92 amino acids, and luteinizing hormone (LH) and human chorionic gonadotropin (HCG) The subunits are very similar, and the beta subunit consists of 115 amino acids and is the building block that determines its specificity. Since FSH is a glycoprotein hormone, and its molecular weight, isoelectric point and other physical and chemical properties are very similar to those of LH and TSH (thyroid-stimulatin...

Claims

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Application Information

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