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CHO cell line expressing porcine beta interferon and application thereof

A technology of beta interferon and cell line, applied in the field of porcine beta interferon and CHO cell line expressing porcine beta interferon, can solve the problems of low product activity, complex production process and product composition, and achieve simple process and product composition. easy-to-control effects

Inactive Publication Date: 2009-12-16
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

The quantification of interferon requires a stable standard product. The vertebrate natural porcine leukocyte interferon in the current quality standards for veterinary biological products promulgated by the Ministry of Agriculture is obtained by inducing porcine leukocytes with chicken Newcastle disease virus. The production process and product components are complex, and the final product Low activity, about 10000AU·mL -1
Compared with other expression systems, vertebrate interferon expressed by mammalian cells can be correctly folded, glycosylated and post-translationally modified, so it is more stable, has higher specific activity, and is more suitable as a standard, especially for IFN-β, while There is currently a lack of standards for vertebrate porcine IFN-β derived from mammalian cells

Method used

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  • CHO cell line expressing porcine beta interferon and application thereof
  • CHO cell line expressing porcine beta interferon and application thereof
  • CHO cell line expressing porcine beta interferon and application thereof

Examples

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Embodiment

[0036] Example. Construction of CHO Expression System for Efficient and Stable Expression of Recombinant Porcine Beta Interferon and Detection of Biological Activity

[0037] 1 Materials and methods

[0038] 1.1 Plasmids, cell lines and virus strains

[0039] PK15 cells (ATCC No.CCL-33) were preserved by our laboratory and cultured in DMEM medium containing 5% FBS. Porcine transmissible gastroenteritis virus (transmissible gastroenteritis virus, TGEV) and pseudorabies virus (pseudorabies virus, PRV) were commercially purchased from the Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, and half of the tissue culture infectious dose was titrated on PK15 cells. 50 , TCID 50 ).

[0040] The stable eukaryotic expression vector pCI-neo (pCN) was purchased from Promega, and neo was expressed under the control of the SV40 early promoter. The CHO-K1 cell line was purchased from ATCC (ATCC No. CCL-61), and the CHO-K1 cells were cultured in DMEM-Ha...

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Abstract

The invention discloses a CHO cell line expressing vertebrate beta interferon, which contains genes coding beta interferon of the animal. The invention particularly discloses a CHO cell line expressing porcine beta interferon. The invention also relates to a method utilizing the CHO cell line to produce standard plasmids of vertebrate beta interferon and porcine beta interferon obtained by the method, and the porcine beta interferon has higher specific activity and stability, can be used as standard plasmids for porcine beta interferon biological activity detection, can be used for preventing and treating porcine infectious diseases and has significant application prospect and market prospect.

Description

technical field [0001] The present invention relates to a CHO cell line expressing vertebrate interferon beta, which contains a gene encoding said animal interferon beta. More specifically, the present invention discloses a CHO cell line expressing porcine beta interferon. The present invention also relates to a method for producing porcine beta interferon standard products using the above-mentioned CHO cell line and the porcine beta interferon obtained by the method, which has extremely high specific activity and stability and can be used as a biological method for porcine beta interferon. It is used as a standard for activity detection and for the prevention and treatment of porcine infectious diseases. Background technique [0002] Type I interferon is an important part of the natural immune system of vertebrates and is an important research field of modern medicine. Nowadays, the mechanism of evading natural immunity such as interferon when the virus infects the host h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/22C07K14/565C12P21/02A61K38/21A61P31/12
Inventor 步志高陈伟业
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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