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Method for cell perfusion culture to produce recombinant protein by microcarrier technology

A recombinant protein and perfusion culture technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as excessive accumulation of metabolic waste, and achieve the effects of avoiding accumulation, high yield, and increased yield

Active Publication Date: 2013-05-15
哈药集团股份有限公司 +1
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  • Abstract
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AI Technical Summary

Problems solved by technology

Fed-batch culture (Fed-batch), continuous feeding for one harvest, timely supplement of nutrients, but excessive accumulation of metabolic waste

Method used

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  • Method for cell perfusion culture to produce recombinant protein by microcarrier technology
  • Method for cell perfusion culture to produce recombinant protein by microcarrier technology
  • Method for cell perfusion culture to produce recombinant protein by microcarrier technology

Examples

Experimental program
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Effect test

Embodiment

[0047] The CHOK1 used in the present invention (from ATCC number CRL-9618) is a vector recombinant host cell containing a gene encoding erythropoietin (EPO) protein and genetic control elements, and is determined to have high-efficiency expression of the recombinant host cell.

[0048] Once the recombinant host cells are determined and the original cell line is preserved, the technical route of the present invention can be used to expand the culture of recombinant cells expressing erythropoietin (EPO) and harvest the target protein. The steps are as follows:

[0049] 1. Expansion of seed cells

[0050] Recombinant CHOK1 cells expressing erythropoietin (EPO) were revived and amplified step by step through T25 square bottles, T75 square bottles, T150 square bottles, and roller bottles. The culture conditions were 37°C and 5% CO2, and the medium used was DMEM. / F12, add 10% fetal bovine serum as needed, pH 7.2. After the cells grow to about 80% in the spinner bottle, the number...

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Abstract

The invention discloses a method for large-scale and high density animal cell perfusion culture to produce recombinant protein by a microcarrier technology. The method is characterized in that: a shaking bioreactor is employed, a microcarrier suspends in the reactor and is in an uninterrupted flow state, a continuous perfusion way is adopted for continuous supplement of nutrients and metabolic waste is discharged, and at the same time the recombinant protein is harvested.

Description

technical field [0001] The invention belongs to the field of biopharmaceutical technology. Specifically, it relates to a method for producing protein by using microcarrier technology to perfuse and culture animal cells. Background technique [0002] As therapeutic drugs, recombinant protein drugs have increasingly highlighted their important medical and commercial value. In general, larger, more complex proteins are produced using mammalian cell expression systems. Therefore, the high-level production of recombinant protein drugs depends on the selection and optimization of the culture process of mammalian cells. [0003] At present, there are many animal cell culture techniques, which can be summarized into three categories, namely adherent culture, suspension culture and immobilized culture. [0004] Adherent culture means that cells are attached to a certain solid surface for monolayer culture. Since the cells are fixed on the surface, no complex cell shut-off device...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00C12R1/91
Inventor 孟庆勇张秀芹张雪亭刘铁成王延涛李会成吕中华王莹李郑武黄宇红王丽娜
Owner 哈药集团股份有限公司
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