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Growth control of hybridoma cells with an artificially induced EpoR-gp130 heterodimer

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Abstract

IL-6 has been known to modulate the growth of many hybridoma cells and also promote resultant antibody productivity. However, IL-6 is so expensive that the use of IL-6-dependent hybridomas for industrial antibody production is not practical. In this study, we aimed at designing antibody/gp130 and antibody/EpoR chimeras which could tightly control cell growth in response to more affordable cognate antigen. Retroviral vectors encoding VH or VL region of anti-hen egg lysozyme (HEL) antibody HyHEL-10 tethered to a pair of extracellular D2/transmembrane domains of erythropoietin receptor (EpoR) and cytoplasmic domains of either EpoR or gp130, were constructed, and a homodimeric or a heterodimeric pair of chimeric receptor combinations (VH-gp130 and VL-gp130 or VH-gp130 and VL-EpoR) were expressed in an IL-6-dependent hybridoma 7TD1. The chimeric receptor-derived growth signal was observed in both combinations, while some residual growth signal was observed in the absence of HEL. To reduce interchain interaction between the two receptor chains, we introduced mutations to the transmembrane domain of both chimera combinations. Consequently, the heterodimeric combination of VH-gp130 and VL-EpoR showed clear HEL-dependent cell growth, while the homodimeric combination of VH-gp130 and VL-gp130 showed reduced cell growth in the absence of HEL. This is the first report that an EpoR-gp130 cytoplasmic domain heterodimer could transduce a growth signal in hybridoma cells, indicating tight and economical growth control of hybridoma cells via our chimeric receptors.

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Acknowledgments

We are grateful to Dr. M. Hibi (Osaka University) for human gp130 expression vector, Dr. T. Kitamura (The University of Tokyo) for retroviral expression system. This work was supported by Grants-in-Aid for Scientific Research (S13854003 and 16023217) from the MEXT, Japan.

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Authors and Affiliations

  1. Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan

    Masahiro Kawahara, Yuko Ogo, Hiroshi Ueda & Teruyuki Nagamune

  2. Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, 301, Bioscience Building, 5-1-5, Kashiwa-no-ha, Kashiwa, 277-8562, Japan

    Kouhei Tsumoto

  3. Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University, Aoba, Aramaki, Aoba-ku, Sendai, 980-8579, Japan

    Izumi Kumagai

Authors
  1. Masahiro Kawahara
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  2. Yuko Ogo
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  3. Kouhei Tsumoto
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  4. Izumi Kumagai
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  5. Hiroshi Ueda
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  6. Teruyuki Nagamune
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Correspondence to Masahiro Kawahara.

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Kawahara, M., Ogo, Y., Tsumoto, K. et al. Growth control of hybridoma cells with an artificially induced EpoR-gp130 heterodimer. Cytotechnology 52, 171–179 (2006). https://doi.org/10.1007/s10616-006-9035-2

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  • DOI: https://doi.org/10.1007/s10616-006-9035-2

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