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Effects of autophagy inducers on recombinant antibody production in insect cells

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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.

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References

  • Blommaart EF, Luiken JJ, Blommaart PJ, van Woerkom GM, Meijer AJ (1995) Phosphorylation of ribosomal protein S6 is inhibitory for autophagy in isolated rat hepatocytes. J Biol Chem 270:2320–2326

    Article  CAS  Google Scholar 

  • Butler M, Meneses-Acosta A (2012) Recent advances in technology supporting biopharmaceutical production from mammalian cells. Appl Microbiol Biotechnol 96:885–894

    Article  CAS  Google Scholar 

  • Cerni S, Shafer D, To K, Venketaraman V (2019) Investigating the role of everolimus in mTOR inhibition and autophagy promotion as a potential host-directed therapeutic target in mycobacterium tuberculosis infection. J Clin Med 8:232

    Article  CAS  Google Scholar 

  • Chames P, van Regenmortel M, Weiss E, Baty D (2009) Therapeutic antibodies: successes, limitations and hopes for the future. Br J Pharmacol 157:220–233

    Article  CAS  Google Scholar 

  • Chen Q, Kang J, Fu C (2018) The independence of and associations among apoptosis, autophagy, and necrosis. Signal Transduct Target Ther 3:18. doi: https://doi.org/10.1038/s41392-018-0018-5. (eCollection)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cho CS, Chang Z, Elkahwaji J, Scheunemann TL, Manthei ER, Colburn M, Knechtle SJ, Hamawy MM (2003) Rapamycin antagonizes cyclosporin A- and tacrolimus (FK506)-mediated augmentation of linker for activation of T cell expression in T cells. Int Immunol 15:1369–1378

    Article  CAS  Google Scholar 

  • Das G, Shravage BV, Baehrecke EH (2012) Regulation and function of autophagy during cell survival and cell death. Cold Spring Harb Perspect Biol 4:a008813

    Article  Google Scholar 

  • Ding L, Nan WH, Zhu XB, Li XM, Zhou LY, Chen HJ, Yu L, Khan FU, Zhong HB, Shi XJ (2019) Rapamycin and FK506 derivative TH2849 could ameliorate neurodegenerative diseases through autophagy with low immunosuppressive effect. CNS Neurosci Ther 25:452–464

    Article  CAS  Google Scholar 

  • Felberbaum RS (2015) The baculovirus expression vector system: A commercial manufacturing platform for viral vaccines and gene therapy vectors. Biotechnol J 10:702–714

    Article  CAS  Google Scholar 

  • Ha TK, Kim YG, Lee GM (2014) Effect of lithium chloride on the production and sialylation of Fc-fusion protein in Chinese hamster ovary cell culture. Appl Microbiol Biotechnol 98:9239–9248

    Article  CAS  Google Scholar 

  • Ha J, Guan KL, Kim J (2015) AMPK and autophagy in glucose/glycogen metabolism. Mol Aspects Med 46:46–62

    Article  CAS  Google Scholar 

  • Hausch F, Kozany C, Theodoropoulou M, Fabian AK (2013) FKBPs and the Akt/mTOR pathway. Cell Cycle 12:2366–2370

    Article  CAS  Google Scholar 

  • Kim CL, Ha TK, Lee GM (2016) Combinatorial treatment with lithium chloride enhances recombinant antibody production in transiently transfected CHO and HEK293E cells. Biotechnol Bioproc Eng 21:667–675

    Article  CAS  Google Scholar 

  • Kim D, Hwang HY, Kim JY, Lee JY, Yoo JS, Marko-Varga G, Kwon HJ (2017) FK506, an immunosuppressive drug, induces autophagy by binding to the V-ATPase catalytic subunit A in neuronal cells. J Proteome Res 16:55–64

    Article  CAS  Google Scholar 

  • Kunert R, Reinhart D (2016) Advances in recombinant antibody manufacturing. Appl Microbiol Biotechnol 100:3451–3461

    Article  CAS  Google Scholar 

  • Lai Y, Suo S, Wang R, Kong X, Hu Y, Tang D, Shi H, Chen S, Hu H (2018) Trends involving monoclonal antibody (mAb) research and commercialization: a scientometric analysis of IMS Lifecyle R&D Focus Database (1980–2016). Hum Vaccin Immunother 14:847–855

    Article  Google Scholar 

  • Lee JS, Lee GM (2012) Rapamycin treatment inhibits CHO cell death in a serum-free suspension culture by autophagy induction. Biotechnol Bioeng 12:3093–3102

    Article  Google Scholar 

  • Luckow VA (1995) Protein production and processing from baculovirus expression vectors. In: Shuler ML, Wood HA, Granados RR, Hammer DA (eds) Baculovirus expression systems and biopesticides. Wiley-Liss, New York, pp 51–90

    Google Scholar 

  • Mao CD, Hoang P, DiCorleto PE (2001) Lithium inhibits cell cycle progression and induces stabilization of p53 in bovine aortic endothelial cells. J Biol Chem 276:26180–26188

    Article  CAS  Google Scholar 

  • Moruno F, Pérez-Jiménez E, Knecht E (2012) Regulation of autophagy by glucose in mammalian cells. Cells 1:372–395

    Article  CAS  Google Scholar 

  • Motoi Y, Shimada K, Ishiguro K, Hattori N (2014) Lithium and autophagy. ACS Chem Neurosci 5:434–442

    Article  CAS  Google Scholar 

  • Nakagaki T, Satoh K, Ishibashi D, Fuse T, Sano K, Kamatari YO, Kuwata K, Shigematsu K, Iwamaru Y, Takenouchi T, Kitani H, Nishida N, Atarashi R (2013) FK506 reduces abnormal prion protein through the activation of autolysosomal degradation and prolongs survival in prion-infected mice. Autophagy 9:1386–1394

    Article  CAS  Google Scholar 

  • Neufeld TP (2012) Autophagy and cell growth–the yin and yang of nutrient responses. J Cell Sci 125:2359–2368

    CAS  PubMed  PubMed Central  Google Scholar 

  • Ohmuro-Matsuyama Y, Katsuda T, Yamaji H (2019) Effects of lithium on the secretory production of recombinant antibody from insect cells. In Vitro Cell Dev Biol Anim 55:1–6

    Article  CAS  Google Scholar 

  • Pengo N, Scolari M, Oliva L, Milan E, Mainoldi F, Raimondi A, Fagioli C, Merlini A, Mariani E, Pasqualetto E, Orfanelli U, Ponzoni M, Sitia R, Casola S, Cenci S (2013) Plasma cells require autophagy for sustainable immunoglobulin production. Nat Immunol 14:298–305

    Article  CAS  Google Scholar 

  • Sonoda H, Kumada Y, Katsuda T, Yamaji H (2012) Production of single-chain Fv-Fc fusion protein in stably transformed insect cells. Biochem Eng J 67:77–83

    Article  CAS  Google Scholar 

  • Walsh G (2006) Biopharmaceutical benchmarks 2006. Nat Biotechnol 24:769–776

    Article  CAS  Google Scholar 

  • Weiner GJ (2015) Building better monoclonal antibody-based therapeutics. Nat Rev Cancer 15:361–370

    Article  CAS  Google Scholar 

  • Yamaji H (2014) Suitability and perspectives on using recombinant insect cells for the production of virus-like particles. Appl Microbiol Biotechnol 98:1963–1970

    Article  CAS  Google Scholar 

Download references

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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

    Ryou Nakanuma, Kyoko Masumi-Koizumi, Yuki Ohmuro-Matsuyama, Tomohisa Katsuda & Hideki Yamaji

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  1. Ryou Nakanuma
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  2. Kyoko Masumi-Koizumi
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  3. Yuki Ohmuro-Matsuyama
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  4. Tomohisa Katsuda
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  5. Hideki Yamaji
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Correspondence to Hideki Yamaji.

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Nakanuma, R., Masumi-Koizumi, K., Ohmuro-Matsuyama, Y. et al. Effects of autophagy inducers on recombinant antibody production in insect cells. Cytotechnology 73, 299–305 (2021). https://doi.org/10.1007/s10616-020-00423-6

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  • DOI: https://doi.org/10.1007/s10616-020-00423-6

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