Effects of autophagy inducers on recombinant antibody production in insect cells
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ORIGINAL PAPER
Effects of autophagy inducers on recombinant antibody production in insect cells Ryou Nakanuma . Kyoko Masumi-Koizumi . Yuki Ohmuro-Matsuyama . Tomohisa Katsuda . Hideki Yamaji
Received: 25 March 2020 / Accepted: 18 September 2020 Ó Springer Nature B.V. 2020
Abstract Insect cells have recently proven to be an excellent platform for the high-level production of functional recombinant proteins. Autophagy is an important mechanism that promotes cell survival by eliminating damaged organelles and protein aggregates, and it also may influence recombinant protein production. In the present study, we compared the effects that autophagy inducers rapamycin, everolimus, and lithium chloride exert on recombinant lepidopteran insect cells that secrete an engineered antibody molecule. Compared with nontreatment, treatment with either rapamycin or everolimus prolonged cell growth to allow high cell density, improved viability in the declining phase, and then increased the yield of secreted antibodies. These positive effects appeared to be induced via autophagy since autophagosomes were clearly detected, particularly in cells treated with rapamycin or everolimus. Unlike rapamycin, another autophagy inducer, FK506, was ineffective in insect cells. The addition of an appropriate autophagy inducer may be effective in increasing the productivity of recombinant proteins in insect cells.
R. Nakanuma K. Masumi-Koizumi Y. Ohmuro-Matsuyama T. Katsuda H. Yamaji (&) Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan e-mail: [email protected]
Keywords Insect cell culture Recombinant protein production Autophagy inducer Rapamycin Everolimus Lithium chloride
Introduction In recent decades, various biologics such as therapeutic antibodies have been developed and approved (Walsh 2006; Weiner 2015; Lai et al. 2018). Such biologics are generally produced as recombinant proteins using cell culture technology and recombinant DNA technology. Mammalian cells such as Chinese hamster ovary (CHO) cells and NS0 cells are major host cells for the production of recombinant proteins that are used as biologics (Butler et al. 2012; Kunert et al. 2016), but mammalian cells have disadvantages such as complicated culture operations and high production costs (Chames et al. 2009). Insect cells have recently been used as an excellent platform for the high-level production of functional recombinant proteins (Felberbaum 2015). Compared with mammalian cells, insect cells are easier and more costeffective to cultivate, and they can be grown to high cell density in suspension with a serum-free medium. Insect cells can also produce significant amounts of recombinant proteins through post-translational processing and modifications that are similar to those
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Cytotechnology
performed in mammalian cells (Luckow 1995; Yamaji 2014). Many recent reports have detailed the influences that autophagy exe
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