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Electrostatic engineering of the interface between heavy and light chains promotes antibody Fab fragment production

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Abstract

Monoclonal antibodies and antibody fragments are used for diverse diagnostic and therapeutic applications. We have investigated the secretory production of Fab fragments from insect cells cotransfected with plasmid vectors carrying heavy- and light-chain genes. In the present study, to promote the formation of the disulfide bond between the heavy and light chains, some positively charged amino acid residues were introduced near the cysteine residue for the disulfide bond at the C-terminus of CL, while some negatively charged amino acid residues were added near the cysteine residue for the disulfide bond at the C-terminus of CH1. This electrostatic steering led to an increase in Fab secretions from insect cells.

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Acknowledgments

The authors thank Dr. Y. Kumada of the Kyoto Institute of Technology for providing us the plasmids encoding the Hc and Lc genes of the 3A21 Fab fragment. This research was partially supported by the programs for developing key technologies for discovering and manufacturing pharmaceuticals used for next-generation treatments and diagnoses from both the Ministry of Economy, Trade and Industry, Japan (METI) and the Japan Agency for Medical Research and Development (AMED).

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Authors and Affiliations

  1. Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan

    Yuki Ohmuro-Matsuyama, Keita Mori, Hirotsugu Hamada & Hideki Yamaji

  2. Manufacturing Technology Association of Biologics (MAB), c/o Center for Business Incubation, Kobe University, 1-5-6 Minatojima-Minamimachi, Chuo-ku, Kobe, 650-0047, Japan

    Yuki Ohmuro-Matsuyama & Hideki Yamaji

  3. Chemical Resources Laboratory, Tokyo Institute of Technology, R1-18, 4259 Nagatsutacho, Midori-ku, Yokohama, 226-8503, Japan

    Hiroshi Ueda

Authors
  1. Yuki Ohmuro-Matsuyama
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  2. Keita Mori
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  3. Hirotsugu Hamada
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  4. Hiroshi Ueda
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  5. Hideki Yamaji
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Correspondence to Hideki Yamaji.

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Ohmuro-Matsuyama, Y., Mori, K., Hamada, H. et al. Electrostatic engineering of the interface between heavy and light chains promotes antibody Fab fragment production. Cytotechnology 69, 469–475 (2017). https://doi.org/10.1007/s10616-016-9955-4

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  • DOI: https://doi.org/10.1007/s10616-016-9955-4

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