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A kind of purification method of recombinant human follicle-stimulating hormone

A technology of human follicle-stimulating hormone and purification method, applied in the fields of peptide preparation method, chemical instrument and method, organic chemistry, etc., can solve the problems of complex purification, many control points, difficult controllability of purification process, etc. Simple, easy-to-scale effects

Active Publication Date: 2015-11-18
QILU PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These two patents involve many purification process steps, the purification is relatively complicated, and there are many control points in the industrial production process, which brings certain difficulties to the controllability of the purification process; in addition, the reverse phase chromatography process is relatively slow and requires In the end, the product needs to be tested for organic residues, and the storage of organic solvents must be considered when it is used in pharmaceutical production in the future, which brings some inconvenience to pharmaceutical production.

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  • A kind of purification method of recombinant human follicle-stimulating hormone
  • A kind of purification method of recombinant human follicle-stimulating hormone

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Embodiment 1

[0030] A method for purifying recombinant human follicle-stimulating hormone, the specific steps of which are:

[0031] Dye affinity chromatography

[0032] (1) Equilibrate the BlueSepharose CL-6B chromatography column (1.6×10cm) with 200ml of Tris-HCl buffer solution with a concentration of 100mmol / L and a pH of 8.0, and filter the clarified and filtered cell culture solution containing the target protein of recombinant human follicle-stimulating hormone Sample loading, so that the recombinant human follicle-stimulating hormone target protein is adsorbed on the dye affinity chromatography medium;

[0033] (2) Use Tris-HCl buffer containing 2.0mol / L NaCl at a concentration of 100mmol / L and a pH of 8.0 as the eluent to elute the column treated in step (1), so that the target protein passes through the dye affinity and a chromatographic column to collect the eluted protein solution;

[0034] Hydrophobic Interaction Chromatography

[0035] (3) Equilibrate Phenyl Sepharose 6 (h...

Embodiment 2

[0042] A method for purifying recombinant human follicle-stimulating hormone, the specific steps of which are:

[0043] Dye affinity chromatography

[0044] (1) Equilibrate the BlueSepharose High Performance chromatography column (1.6×10cm) with 200ml of Tris-HCl buffer solution with a concentration of 50mmol / L and a pH of 7.0, and filter the clarified and filtered cell culture supernatant containing the target protein of recombinant human follicle-stimulating hormone. Sample loading, so that the recombinant human follicle-stimulating hormone target protein is adsorbed on the dye affinity chromatography medium;

[0045] (2) Use Tris-HCl buffer containing 1.0mol / L NaCl at a concentration of 50mmol / L and a pH of 7.0 as the eluent to elute the column treated in step (1) so that the target protein passes through the dye affinity and a chromatographic column to collect the eluted protein solution;

[0046] Hydrophobic Interaction Chromatography

[0047] (3) Equilibrate Phenyl Se...

Embodiment 3

[0054] A method for purifying recombinant human follicle-stimulating hormone, the specific steps of which are:

[0055] Dye affinity chromatography

[0056] (1) Equilibrate the BlueSepharose6FastFlow chromatography column (1.6×10cm) with 200ml of phosphate buffer solution with a concentration of 20mmol / L and a pH of 10.0, and supernatant the clarified and filtered cell culture supernatant containing the target protein of recombinant human follicle-stimulating hormone In this way, the recombinant human follicle-stimulating hormone target protein is adsorbed on the dye affinity chromatography medium;

[0057] (2) Use a phosphate buffer solution containing 2.5mol / L NaCl at a concentration of 20mmol / L and a pH of 10.0 as the eluent to elute the chromatography column treated in step (1) to allow the target protein to pass through the dye affinity Chromatographic column, collecting the eluted protein solution;

[0058] Hydrophobic Interaction Chromatography

[0059] (3) Equilibra...

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Abstract

The invention relates to the production of genetically engineered drugs by using a recombinant DNA (deoxyribonucleic acid) technique and belongs to the field of biotechnology. The invention particularly relates to a method for purifying recombinant human follicle stimulating hormone. According to the method, a high-quality recombinant human follicle stimulating hormone interest protein is acquired by sequentially treating cellular supernatant containing the recombinant human follicle stimulating hormone through dye affinity chromatography, hydrophobic interaction chromatography, desalination chromatography and hydroxyapatite chromatography. The method provided by the invention has the characteristics that the technique is simple, the cost is low, the target product is high in purity, the middle process is convenient to control, and the production is easy to amplify in a large scale.

Description

technical field [0001] The invention relates to the production of genetic engineering drugs by using recombinant DNA technology, in particular to a method for purifying recombinant human follicle-stimulating hormone, which belongs to the field of biotechnology. Background technique [0002] Follicle Stimulating Hormone (FSH), also known as follicle-stimulating hormone, is an injectable protein belonging to the gonadotropin class. Naturally synthesized and secreted by pituitary basophils, FSH can be recombinantly produced for medicinal purposes, or can be isolated from the urine of postmenopausal women. FSH can be used for the treatment of infertility and reproductive disorders in female and male patients. At present, there is urinary follicle-stimulating hormone for injection extracted from urine on the market in China, and the trade name is Lishenbao; products produced by genetic recombination technology have also been released. Imported and marketed, its purity is higher,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/59C07K1/22C07K1/20C07K1/16
CPCC07K14/59
Inventor 陈俊良孙丽霞王晶翼孙波邢婷婷孙旭张宗辉
Owner QILU PHARMA CO LTD