Abstract
In our previous study, we developed a novel cycloheximide resistance (cyh R) marker for use in mammalian cells. In experiments using human monoclonal antibody genes, vectors containing this cyh R marker produced cycloheximide resistant CHO clones whose maximum expression levels were significantly higher than that of the neo R/G418 clones. The expression levels of cyh R clones were 3 to 4 times higher than those of neo R clones, in protein-free suspension culture. The maximum yield of antibody in cyh R clones reached over 700 mg/L in a typical fed-batch culture using a 3L-bioreactor. Cell growth studies showed no difference in doubling times between cyh R and neo R clones expressing antibodies and stability of expression in the absence of antibiotic was also demonstrated. The cyh R system thus provides a significantly more efficient method for selection of high-expressing CHO clones without the need for gene amplification, and in a substantially shorter time frame than methods currently in use.
Keywords
- Human Monoclonal Antibody
- Antibody Concentration
- Maximum Expression Level
- Antibody Expression
- Dominant Selectable Marker
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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References
Misawa E, Yajima H and Kondo K: A novel selectable marker for increased recombinant protein expression in transfected mammalian cells. (Submitted)
Ishida I: The KM Mouse™; For efficient and rapid production of high affinity human monoclonal antibodies, 12th Annual International Conference on Antibody Engineering, San Diego. (2001)
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© 2003 Springer Science+Business Media Dordrecht
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Tsukahara, M. et al. (2003). High Expression of Human Monoclonal Antibodies in CHO Cells Selected Using a Cycloheximide Resistance Marker. In: Yagasaki, K., Miura, Y., Hatori, M., Nomura, Y. (eds) Animal Cell Technology: Basic & Applied Aspects. Animal Cell Technology: Basic & Applied Aspects, vol 13. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-0726-8_20
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DOI: https://doi.org/10.1007/978-94-017-0726-8_20
Publisher Name: Springer, Dordrecht
Print ISBN: 978-90-481-6557-5
Online ISBN: 978-94-017-0726-8
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