Journal of Microbiology and Biotechnology
The Korean Society for Microbiology and Biotechnology publishes the Journal of Microbiology and Biotechnology.

2019 ; Vol.29-1: 55~58

AuthorJuyoung Byun, Sena Yoon, Yunji Jeong, Uitaek Oh, Sujin Cho, Jeongsoo Park, Yongsu Jeong, Kwanghee Baek, Jaeseung Yoon
Place of dutyPanGen Biotech Inc, Republic of Korea
TitleEfficient Development of Stable Recombinant Chinese Hamster Ovary (rCHO) Cell Lines to Produce Antibodies by Using Dimethyl Sulfoxide (DMSO) in Electroporation
PublicationInfo J. Microbiol. Biotechnol.2019 ; Vol.29-1
AbstractDevelopment of stable rCHO cell lines is still time consuming and labor intensive, although it is a critical step in the commercial development of recombinant antibodies. The current work demonstrates, for the first time, that electroporation of CHO cells with DMSO can enhance stable expression of recombinant antibodies in rCHO cells. Electroporation with DMSO resulted in an average 3.7-fold and 2.8-fold increases in expression levels of aflibercept and pembrolizumab, respectively, in pools of stable rCHO cells. It also resulted in an average of 2.2-fold and 2.6-fold increases in the expression of aflibercept and pembrolizumab, respectively, in single-cell derived rCHO clones. Simple batch cultures of rCHO cell clones with the highest expression produced 1.0 g/l for aflibercept and 1.4 g/l for pembrolizumab without a time-consuming gene amplification process. Electroporation with DMSO also shortened the development of rCHO cell lines to 2-3 months, allowing rapid establishment of stable rCHO cell lines with a desirable expression level antibodies.
Key_wordDimethyl sulfoxide, stable cell line, chinese hamster ovary cell, recombinant antibody, electroporation
  1. Jesus MD, Wurm FM. 2011. Manufacturing recombinant proteins in kg-ton quantities using animal cells in bioreactors. Eur. J. Pharm. Biopharm. 78: 184-188.
    Pubmed CrossRef
  2. Durocher Y, Butler, M. 2009. Expression systems for therapeutic glycoprotein production. Curr. Opin. Biotechnol. 20: 700-707.
    Pubmed CrossRef
  3. Dietmair S, Nielsen LK, Timmins NE. 2010. Engineering a mammalian super producer. J. Chem. Technol. Biotechnol. 86:905-914.
  4. Noh SM, Sathyamurthy M, Lee GM. 2013. Development of recombinant Chinese hamster ovary cell lines for therapeutic protein production. Curr Opin. Chem. Eng. 2: 391-397.
  5. Baldi L, Hacker DL, Adam M, Wurm FM. 2007. Recombinant protein production by large-scale transient gene expression in mammalian cells: state of art and future perspectives. Biotechnol. Lett. 29: 677-684.
    Pubmed CrossRef
  6. Bertschinger M, Schertenleib A, Cavey J, Hacker DL, Wurm FM. 2008. The kinetics of polyethylenimine-mediated transfection in suspension cultures of Chinese hamster ovary cells. Mol Biotechnol. 40: 136-143.
    Pubmed CrossRef
  7. Steger K, Brady J, Wang W, Duskin M, Donato K, Peshwa M. 2015. CHO-S antibody titers >1 gram/liter using flow electroporation-mediated gene expression followed by rapid migration to high-yield stable cell lines. J. Biomol. Screen. 20:545-551.
    Pubmed CrossRef Pubmed Central
  8. Cervera L, Fuenmayer J, Gonzalez-Dominguez I, GutierrezGranados S, Segura MM, Godia F. 2015. Selection and optimization enhancer additives for increased virus-like particle production in HEK293 suspension cell cultures. Appl. Microbiol. Biotechnol. 99: 9935-9949.
    Pubmed CrossRef
  9. Melkonyan H, Sorg C, Klempt M. 1996. Electroporation efficiency in mammalian cells is increased by dimethyl sulfoxide (DMSO). Nuc. Acids Res. 24: 4356-4357.
    Pubmed CrossRef Pubmed Central
  10. Ye J, Kober V, Tellers M, Naji Z, Salmon P, Markusen JF. 2009. High-level protein expression in scalable CHO transient transfection. Biotechnol. Bioeng. 103: 542-551.
    Pubmed CrossRef
  11. Daly TJ, Fandl JP, Papadopoulos NJ. 2006. Fusion protein capable of binding VEGF. US Patent publication No 7087411B2.
  12. Yoon J, Baek K, Byun T, Park J. 2014. Expression vector animal cells. US Patent Publication No 20140038233.
  13. Ciudad CJ, Urlaub G, Chasin LA. 1988. Deletion analysis of the Chinese hamster dihydrofolate reductase gene promoter. J. Biol. Chem. 263: 16274-16282.
  14. Paredes V, Park JS, Jeong Y, Yoon J, Baek K. 2013. Unstable expression of recombinant antibody during long-term culture of CHO cells is accompanied by histone H3 hypoacetylation. Biotechnol. Lett. 35: 987-993.
    Pubmed CrossRef

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