Overexpression of mutant cell division cycle 25 homolog B (CDC25B) enhances the efficiency of selection in Chinese hamster ovary cells

Cytotechnology - Tập 65 - Trang 1017-1026 - 2013
Kyoung Ho Lee1, Tomomi Tsutsui2, Kohsuke Honda1, Hisao Ohtake1, Takeshi Omasa1,2
1Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Japan
2Institute of Technology and Science, The University of Tokushima, Tokushima, Japan

Tóm tắt

The effects of mutant cell division cycle 25 homolog B (CDC25B) overexpression on the generation of cells producing a monoclonal antibody were investigated in Chinese hamster ovary (CHO) cells. Mutant CDC25B (m-CDC25B) expression plasmids were transfected into CHO DG44-derived cells producing a monoclonal antibody, and the frequency of highly producing cells was assessed following gene amplification in the presence of 250 nM methotrexate. Most of the clones obtained from the m-CDC25B-overexpressing cells had higher antibody titers than did mock-transfected control cells. This arose from either higher transgene copy numbers or higher mRNA expression levels for the antibody. However, the high mRNA expression levels were not always accompanied by increases in transgene copy numbers. Our results suggest that cells producing high levels of a monoclonal antibody can be selected efficiently using m-CDC25B overexpression.

Tài liệu tham khảo

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