Establishment of a novel cell line, CHO-MK, derived from Chinese hamster ovary tissues for biologics manufacturing

Kenji Masuda; Michi Kubota; Yuto Nakazawa; Chigusa Iwama; Kazuhiko Watanabe; Naoto Ishikawa; Yumiko Tanabe; Satoru Kono; Hiroki Tanemura; Shinichi Takahashi; Tomohiro Makino; Takeshi Okumura; Takayuki Horiuchi; Koichi Nonaka; Sei Murakami; Masamichi Kamihira; Takeshi Omasa

doi:10.1016/j.jbiosc.2024.02.005

Establishment of a novel cell line, CHO-MK, derived from Chinese hamster ovary tissues for biologics manufacturing

J Biosci Bioeng. 2024 Mar 11:S1389-1723(24)00057-4. doi: 10.1016/j.jbiosc.2024.02.005. Online ahead of print.

Authors

Kenji Masuda¹, Michi Kubota², Yuto Nakazawa³, Chigusa Iwama², Kazuhiko Watanabe³, Naoto Ishikawa³, Yumiko Tanabe³, Satoru Kono³, Hiroki Tanemura³, Shinichi Takahashi³, Tomohiro Makino³, Takeshi Okumura¹, Takayuki Horiuchi⁴, Koichi Nonaka¹, Sei Murakami⁵, Masamichi Kamihira⁶, Takeshi Omasa⁷

Affiliations

¹ Biologics Division, Biologics Technology Research Laboratories I, Daiichi Sankyo Co., Ltd., 2716-1 Kurakake, Akaiwa, Chiyoda-machi, Gunma 370-0503, Japan; Manufacturing Technology Association of Biologics, 2-6-16 Shinkawa, Chuo-ku, Tokyo 104-0033, Japan.
² Manufacturing Technology Association of Biologics, 2-6-16 Shinkawa, Chuo-ku, Tokyo 104-0033, Japan; Chitose Laboratory Corp., KSP EAST511, 3-2-1 Sakado, Takatsu-ku, Kawasaki, Kanagawa 213-0012, Japan.
³ Biologics Division, Biologics Technology Research Laboratories I, Daiichi Sankyo Co., Ltd., 2716-1 Kurakake, Akaiwa, Chiyoda-machi, Gunma 370-0503, Japan.
⁴ Chitose Laboratory Corp., KSP EAST511, 3-2-1 Sakado, Takatsu-ku, Kawasaki, Kanagawa 213-0012, Japan.
⁵ Manufacturing Technology Association of Biologics, 2-6-16 Shinkawa, Chuo-ku, Tokyo 104-0033, Japan.
⁶ Manufacturing Technology Association of Biologics, 2-6-16 Shinkawa, Chuo-ku, Tokyo 104-0033, Japan; Department of Chemical Engineering, Faculty of Engineering, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan.
⁷ Manufacturing Technology Association of Biologics, 2-6-16 Shinkawa, Chuo-ku, Tokyo 104-0033, Japan; Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: omasa@bio.eng.osaka-u.ac.jp.

PMID: 38472071
DOI: 10.1016/j.jbiosc.2024.02.005

Abstract

Chinese hamster ovary (CHO) cells are widely used as a host for producing recombinant therapeutic proteins due to advantages such as human-like post-translational modification, correct protein folding, higher productivity, and a proven track record in biopharmaceutical development. Much effort has been made to improve the process of recombinant protein production, in terms of its yield and productivity, using conventional CHO cell lines. However, to the best of our knowledge, no attempts have been made to acquire new CHO cell lines from Chinese hamster ovary. In this study, we established and characterized a novel CHO cell line, named CHO-MK, derived from freshly isolated Chinese hamster ovary tissues. Some immortalized cell lines were established via sub-culture derived from primary culture, one of which was selected for further development toward a unique expression system design. After adapting serum-free and suspension culture conditions, the resulting cell line exhibited a considerably shorter doubling time (approximately 10 h) than conventional CHO cell lines (approximately 20 h). Model monoclonal antibody (IgG₁)-producing cells were generated, and the IgG₁ concentration of fed-batch culture reached approximately 5 g/L on day 8 in a 200-L bioreactor. The cell bank of CHO-MK cells was prepared as a new host and assessed for contamination by adventitious agents, with the results indicating that it was free from any such contaminants, including infectious viruses. Taking these findings together, this study showed the potential of CHO-MK cells with a shorter doubling time/process time and enhanced productivity in biologics manufacturing.

Keywords: CHO expression system; CHO-MK; Cell line development; Cell line establishment; Chinese hamster ovary cell line; Monoclonal antibody manufacturing; Serum-free and suspension culture adaptation.