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Cellular expression system for stably expressing anti-TNF-alpha monoclonal antibody

A monoclonal antibody and stable expression technology, applied in the field of biomedicine, can solve the problems of high treatment cost, poor genetic stability, dependence on imports, etc., and achieve the effect of improving the proliferation speed and antibody production performance, and improving genetic stability

Active Publication Date: 2017-10-10
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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Problems solved by technology

However, the domestic production of this drug is still blank, and the clinical practice is all dependent on imports, and the treatment cost is high
[0004] In addition, the currently commercially available anti-TNF-α monoclonal antibody co-expresses the light chain and heavy chain through two plasmids, and the genetic stability is relatively poor

Method used

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  • Cellular expression system for stably expressing anti-TNF-alpha monoclonal antibody
  • Cellular expression system for stably expressing anti-TNF-alpha monoclonal antibody
  • Cellular expression system for stably expressing anti-TNF-alpha monoclonal antibody

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Embodiment Construction

[0027] The present invention is described in further detail below by specific embodiment, and following embodiment is descriptive only, is not limiting, can not limit protection scope of the present invention with this.

[0028] The present invention provides a kind of cell expression system stably expressing anti-TNF-α monoclonal antibody and construction method and application, and specific content comprises:

[0029] A cell expression system for stably expressing anti-TNF-α monoclonal antibody. The codons of the light and heavy chain coding genes optimized according to the cell codon preference are used to construct eukaryotic expression plasmids for the co-expression of the light and heavy chains. On this basis, the Antibody homologous recombination vectors targeting genes (BAK1 and Bax) that do not affect the basic viability of cells and have potential negative effects on cell proliferation and antibody expression and production, and construct CRISPR / Case9 vectors with spe...

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Abstract

The invention relates to a cellular expression system for stably expressing an anti-TNF-alpha monoclonal antibody, and a construction method and application of the cellular expression system. According to the cellular expression system, light and heavy chain encoding genes of the anti-TNF-alpha monoclonal antibody are optimized according to cell codon preference so as to construct anti-TNF-alpha human-mouse chimeric monoclonal antibody homologous recombination vector with a Bak1 gene and the like as a drone, meanwhile, a CRISPR / Case9 vector with a specific cutting effect is constructed for exon regions of drone genes such as Bak1, two vectors are commonly guided into Chinese hamster ovary cells (CHO) by lipofection transfection, and a positive cell strain capable of stably expressing the anti-TNF-alpha monoclonal antibody is obtained through screening. The cellular expression system can be applied to efficient production of the anti-TNF-alpha monoclonal antibody, and the anti-TNF-alpha monoclonal antibody can be used for treating diseases such as rheumatoid arthritis and ankylosing spondylitis.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to the construction and expression of a cell expression system stably expressing anti-TNF-α monoclonal antibody. Background technique [0002] Rheumatoid arthritis (RA) is a chronic systemic immune inflammatory disease of unknown cause, which can cause joint swelling, pain and dysfunction throughout the body. It is one of the main disabling diseases and seriously affects the lives of patients. quality. RA is a worldwide common disease. The prevalence rate in my country is 0.3-0.4%. It can occur at any age, and it is more common in women aged 30-50. [0003] The current drugs for the treatment of RA mainly include nonsteroidal antiinflammatory drugs (NSAIDs), disease-modifying anti-rheumatic drugs (NMARDs), adrenocortical hormones, traditional Chinese medicine preparations and biological preparations, etc. The etiology of RA is complex, involving the effects of cytokines and immunity. The ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/90C12N15/85
CPCC07K16/241C12N15/85C12N15/907C12N2800/22C12N2830/20
Inventor 罗学刚张同存苗峙李潜王鹏赵涧
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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