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Production method for recombinant human granulocyte colony-stimulating factor

A technology of stimulating factor and production method, applied in the field of production of recombinant human granulocyte stimulating factor, can solve the problems of difficulty in controlling ultrasonic power and time, carbonization of protein, inability to completely release recombinant protein, etc., and achieves high speed and large processing capacity. , the effect of reducing difficulty

Active Publication Date: 2013-08-07
BEIJING FOUR RINGS BIOPHARM
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AI Technical Summary

Problems solved by technology

At the same time, it is difficult to control the ultrasonic power and time when using ultrasonic to destroy bacteria. Insufficient ultrasonic can not completely release the recombinant protein, and the ultrasonic time is too long or the power is too high, which will carbonize the protein.

Method used

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Embodiment Construction

[0039] The present invention will be described in further detail below in conjunction with specific embodiments, so as to help understand the content of the present invention.

[0040] The preparation method of the recombinant human granulocyte colony-stimulating factor of the present invention comprises the steps of: a. selecting engineering bacteria pKG931 / HB101 to ferment and cultivate; b. extracting and washing inclusion bodies from the bacterial cells obtained through fermentation and culture to obtain refined inclusion bodies; c. Denaturing the refined inclusion body to obtain a denatured product; d. Refolding the denatured product; e. Performing chromatography on the refolded product to obtain recombinant human granulocyte colony-stimulating factor.

[0041] The specific method is:

[0042] a. Select engineering bacteria pKG931 / HB101 provided by Beijing Kangsai Medical Technology Co., Ltd. for fermentation and culture

[0043] Wherein, step a comprises seed expansion c...

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Abstract

The invention discloses production method for a recombinant human granulocyte colony-stimulating factor. According to the invention, an engineered strain of an escherichia coli expressed recombinant human granulocyte colony-stimulating factor is cultured to obtain an inclusion body, the engineered strain of the escherichia coli expressed recombinant human granulocyte colony-stimulating factor is pKG931 / HB101, a culture method comprises multiplication culture and culture in a fermentation cylinder, an antibiotic-free medium containing yeast and peptone is employed in both culture, a high pressure homogenizer is used for breaking of bacteria, and denaturation, renaturation and chromatography are carried out so as to obtain a high purity G-CSF stock solution. The production method provided by the invention has the advantages of a short production period, high production efficiency, a great production scale, especial suitability for industrial production and reduction in production cost.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a production method of recombinant human granulocyte stimulating factor. Background technique [0002] Recombinant human granulocyte-stimulating factor (granulocyte colony-stimulating factor, G-CSF) refers to specifically acting on neutrophil progenitor cells in the hematopoietic system, promoting their proliferation, differentiation into mature neutrophils, and maintaining the survival of cells and their biological A hematopoietic growth factor that functions as a colony-stimulating factor that stimulates bone marrow cell colony formation. It has obvious curative effect on neutrophil increase during bone marrow transplantation, severe neutropenia caused by cancer chemotherapy, and neutropenia accompanied by aplastic anemia. [0003] The biological activity of recombinant G-CSF by genetic engineering technology is similar to that of natural one, and it can be produced in large ...

Claims

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Application Information

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IPC IPC(8): C12P21/02C07K1/18C07K1/14C12R1/19
Inventor 程度胜桑建彬韩明娣龙应国王晓建
Owner BEIJING FOUR RINGS BIOPHARM
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