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Fucose content of monoclonal antibodies can be controlled by culture medium osmolality for high antibody-dependent cellular cytotoxicity

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Abstract

Antibody-dependent cellular cytotoxicity (ADCC) is dependent on the fucose content of oligosaccharides bound to monoclonal antibodies (MAbs). As MAbs with a low fucose content exhibit high ADCC activity, it is important to control the defucosylation levels (deFuc%) of MAbs and to analyze the factors that affect deFuc%. In this study, we observed that the deFuc% was inversely related to culture medium osmolality for MAbs produced in the rat hybridoma cell line YB2/0, with r 2 values as high as 0.92. Moreover, deFuc% exhibited the same correlation irrespective of the type of compound used for regulating osmolality (NaCl, KCl, fucose, fructose, creatine, or mannitol) at a culture scale ranging from 1 to 400 L. We succeeded in controlling MAb deFuc% by maintaining a constant medium osmolality in both perfusion and fed-batch cultures. In agreement with these observations, reverse transcription PCR analyses revealed decreased transcription of genes involved in glycolysis, GDP-fucose supply, and fucose transfer under hypoosmotic conditions.

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Abbreviations

ADCC:

Antibody-dependent cellular cytotoxicity

CHO:

Chinese hamster ovary

dCO2 :

Dissolved carbon dioxide

deFuc:

Defucosylation levels, the extent with which the GlcNAc residue at the reducing terminal is free from glycosylation with fucose

DOE:

Design of experiment

Fut8 :

Alpha-1,6-fucosyltransferase gene

GMD :

GDP-mannose 4,6-dehydratase gene

GlcNAc:

N-acetylglucosamine

MAbs:

Monoclonal antibodies

QbD:

Quality by design

RT-PCR:

Reverse transcription polymerase chain reaction

SPR:

Specific MAb production rate (pg cells−1 d−1)

vvd:

Volume of fresh medium per working volume of reactor per day

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Acknowledgments

We would like to thank Mr. Yuya Kawagoe, Mr. Kazutoshi Maki, Mr. Noriyuki Takahashi, and Ms. Chigusa Hijikata for their expert analyses. We also thank Dr. Mitsuo Sato, Dr. Jun Yamaya, Dr. Kazuhisa Uchida, and Mr. Hiroshi Takasugi for their helpful discussions and encouragement.

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Authors and Affiliations

  1. Bio Process Research and Development Laboratories, Kyowa Hakko Kirin Co., Ltd., 100-1 Hagiwara-machi, Takasaki-shi, Gunma, 370-0013, Japan

    Yoshinobu Konno, Yuki Kobayashi, Ken Takahashi, Eiji Takahashi, Shinji Sakae, Masako Wakitani, Toshiyuki Suzawa & Keiichi Yano

  2. Tokyo Research Park, Kyowa Hakko Kirin Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo, 194-8533, Japan

    Kazuya Yamano

  3. Fuji Research Park, Kyowa Hakko Kirin Co., Ltd., 1188 Shimotogari, Nagaizumi-cho, Suntou-gun, Shizuoka, 411-8731, Japan

    Toshio Ohta

  4. Kyowa Hakko Kirin Pharma, Inc., 212 Carnegie Center, Suite 101, Princeton, NJ, 08540, USA

    Masamichi Koike

  5. Kyowa Hakko Kirin Co., Ltd., 1-6-1 Ohte-machi, Chiyoda-ku, Tokyo, 100-8185, Japan

    Shinji Hosoi

  6. Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu-shi, 376-8515, Japan

    Yoshinobu Konno & Kaori Wakamatsu

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  1. Yoshinobu Konno
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  2. Yuki Kobayashi
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Correspondence to Yoshinobu Konno.

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Konno, Y., Kobayashi, Y., Takahashi, K. et al. Fucose content of monoclonal antibodies can be controlled by culture medium osmolality for high antibody-dependent cellular cytotoxicity. Cytotechnology 64, 249–265 (2012). https://doi.org/10.1007/s10616-011-9377-2

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