Violacein improves recombinant IgG production by controlling the cell cycle of Chinese hamster ovary cells
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ORIGINAL ARTICLE
Violacein improves recombinant IgG production by controlling the cell cycle of Chinese hamster ovary cells Masahide Kido
. Hideaki Idogaki . Kouji Nishikawa . Takeshi Omasa
Received: 16 May 2020 / Accepted: 28 October 2020 Ó Springer Nature B.V. 2020
Abstract Chinese hamster ovary (CHO) cells are used as host cells for industrial monoclonal antibody (mAb) production. Cell cycle control is an effective approach to increase mAb production in the cell culture. Violacein, a purple-colored pigment produced by microorganisms, has diverse bioactive properties and has been proposed for various industrial applications. In this study, we evaluated the potency of violacein for cell cycle control and improvement of recombinant immunoglobulin G (IgG) production in CHO cells. Compared with the control, 0.9 lM violacein in a 14-day fed-batch culture increased the maximum IgG concentration by 37.6% via increasing the specific production rate and cell longevity. Cell cycle analysis showed that violacein induced G1 and G2/M phase arrest. However, the G1 arrest was observed only on day 1, while G2/M arrest lasted more than 3 days, suggesting that G2/M arrest M. Kido (&) H. Idogaki K. Nishikawa Research and Development, OSAKA SODA Co., Ltd, Amagasaki, Hyogo 660-0842, Japan e-mail: [email protected] H. Idogaki e-mail: [email protected] K. Nishikawa e-mail: [email protected] M. Kido T. Omasa Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan e-mail: [email protected]
mediated the violacein-induced enhanced IgG production. Moreover, in line with the increased protein expression, the expression levels of IgG mRNA and nutrient metabolic rates were also increased. N-Linked glycosylation and charge variant profiles were barely affected by violacein treatment. Our results indicate that violacein affects the cell cycle of CHO cells and increases IgG production without changing product quality, showing promise as a mAb production enhancer in CHO cells. The study provides insight into violacein utilization in industrial mAb manufacturing and can help develop advanced, effective mAb production technologies using CHO cell cultures. Keywords CHO cells Violacein Cell culture Cell cycle Recombinant IgG
Introduction Monoclonal antibodies (mAbs) are used therapeutically because of their high target specificity, fewer side effects, and long half-life in the blood. Chinese hamster ovary (CHO) cells are the most widely used host cells for therapeutic antibody production because of their ability to perform appropriate post-translational modifications, such as glycosylation (Xu et al. 2011). Moreover, CHO cells can adapt to serum-free
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Cytotechnology
media and suspension culture, therefore are suitable for large-scale industrial culture. Cell cycle control is a potent way to increase mAb production in CHO cell cultures. There are various approaches to cell cycle control and enhanced protein prod
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