Abstract
Bioreactors are manufactured apparatuses that allow the generation of a specific environment for the highly controlled cultivation of living cells. Originally used for microbial production systems, they have found widespread applications in fields as diverse as vaccine production, plant cell cultivation, and the growth of human brain organoids and exist in equally diverse designs (Chu and Robinson, Curr Opin Biotechnol 12(2):180–187, 2001; Qian et al., Nat Protoc 13:565–580, 2018). Manufacturing of biologics is currently mostly performed using a stirred tank bioreactor and CHO host cells and represents the most “classical” bioreactor production process. In this chapter, we will therefore use the cultivation of suspension Chinese hamster ovary (CHO) cells for recombinant protein production in a stirred tank bioreactor as an example. However, general guidelines provided in this chapter are transferable to different bioreactor types and host cells (Li et al., MAbs 2(5):466–479, 2010).
The preparation and operation of a bioreactor (also referred to as upstream process in a biotechnological/industrial setting) is comprised of three main steps: expansion (generation of biomass), production (batch, fed-batch, or continuous process), and harvest. The expansion of cells can last from few days to weeks depending on the number of cells at the start, the cellular doubling time, and the required biomass to inoculate the production bioreactor. The production phase lasts a few weeks and is a highly sensitive phase as the concentration of different chemicals and physical parameters need to be tightly controlled. Finally, the harvest will allow the separation of the product of interest from large particles and then the desired material (cell culture supernatant or cells) is transferred to the downstream process.
The raw materials used during the upstream phase (all three steps) need to be aligned with the final purpose of the manufactured product, as the presence of residual impurities may have an impact on suitability of the final product for a desired purpose.
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References
Chu L, Robinson DK (2001) Industrial choices for protein production by large-scale cell culture. Curr Opin Biotechnol 12(2):180–187
Li F, Vijayasankaran N, Shen AY, Kiss R, Amanullah A (2010) Cell culture processes for monoclonal antibody production. MAbs 2(5):466–479
Zhu J (2012) Mammalian cell protein expression for biopharmaceutical production. Biotechnol Adv 30(5):1158–1170
Wurm F (2004) Production of recombinant protein therapeutics in cultivated mammalian cells. Nat Biotechnol 22:1393–1398
Kunert R, Reinhart D (2016) Advances in recombinant antibody manufacturing. Appl Microbiol Biotechnol 100:3451–3461
Castan A, Schlutz P, Wenger T, Fischer S (2018) Chapter 7—Cell line development. Biopharmaceuticals processing. ISBN 9780081006238
Hernández Rodríguez T, Frahm B (2020) Design, optimization, and adaptive control of cell culture seed trains. Methods Mol Biol 2095:251–267
van der Valk J, Brunner D, De Smet K, Fex Svenningsen A, Honegger P, Knudsen LE, Lindl T, Noraberg J, Price A, Scarino ML, Gstraunthaler G (2010) Optimization of chemically defined cell culture media—replacing fetal bovine serum in mammalian in vitro methods. Toxicol In Vitro 24(4):1053–1063
Shukla AA, Gottschalk U (2012) Single-use disposable technologies for biopharmaceutical manufacturing. Trends Biotechnol 31(3):147–154
Junne S, Neubauer P (2018) How scalable and suitable are single-use bioreactors? Curr Opin Biotechnol 53:240–247
Xu S, Hoshan L, Jiang R, Gupta B, Brodean E, O’Neill K, Seamans TC, Bowers J, Chen H (2017) A practical approach in bioreactor scale-up and process transfer using a combination of constant P/V and vvm as the criterion. Biotechnol Prog 33(4):1146–1159
Sandadi S, Pedersen H, Bowers JS, Rendeiro D (2011) A comprehensive comparison of mixing, mass transfer, Chinese hamster ovary cell growth, and antibody production using Rushton turbine and marine impellers. Bioprocess Biosyst Eng. 34(7):819–832
Reinhart D, Damjanovic L, Castan A, Ernst W, Kunert R (2018) Differential gene expression of a feed-spiked super-producing CHO cell line. J Biotechnol 285:23–37
Pan X, Streefland M, Dalm C, Wijffels RH, Martens DE (2016) Selection of chemically defined media for CHO cell fed-batch culture processes. Cytotechnology 69(1):39–56
Reinhart D, Damjanovic L, Kaisermayer C, Kunert R (2015) Benchmarking of commercially available CHO cell culture media for antibody production. Appl Microbiol Biotechnol 99(11):4645–4657
Hua J, Erickson LE, Yiin TY, Glasgow LA (1993) A review of the effects of shear and interfacial phenomena on cell viability. Crit Rev Biotechnol 13(4):305–328
Pelton R (2002) A review of antifoam mechanisms in fermentation. J Ind Microbiol Biotechnol 29(4):149–154
Velugula-Yellela SR, Williams A, Trunfio N, Hsu CJ, Chavez B, Yoon S, Agarabi C (2018) Impact of media and antifoam selection on monoclonal antibody production and quality using a high throughput micro-bioreactor system. Biotechnol Prog 34(1):262–270
Demuth C, Varonier J, Jossen V, Eibl R, Eibl D (2016) Novel probes for pH and dissolved oxygen measurements in cultivations from millilitre to benchtop scale. Appl Microbiol Biotechnol 100(9):3853–3863
Maruthamuthu MK, Rudge SR, Ardekani AM, Ladisch MR, Verma MS (2020) Process analytical technologies and data analytics for the manufacture of monoclonal antibodies. Trends Biotechnol 38(10):1169–1186
Xu J, Tang P, Yongky A, Drew B, Borys MC, Liu S, Li ZJ (2019) Systematic development of temperature shift strategies for Chinese hamster ovary cells based on short duration cultures and kinetic modeling. mAbs 11(1):191–204
Odeleye AO, Marsh DT, Osborne MD, Lye GJ, Micheletti M (2014) On the fluid dynamics of a laboratory scale single-use stirred bioreactor. Chem Eng Sci 111(100):299–312
Michl J, Park KC, Swietach P (2019) Evidence-based guidelines for controlling pH in mammalian live-cell culture systems. Commun Biol 2:144
Goldrick S, Lee K, Spencer C, Holmes W, Kuiper M, Turner R, Farid SS (2018) On-line control of glucose concentration in high-yielding mammalian cell cultures enabled through oxygen transfer rate measurements. Biotechnol J 13(4):e1700607
Hoshan L, Jiang R, Moroney J, Bui A, Zhang X, Hang TC, Xu S (2018) Effective bioreactor pH control using only sparging gases. Biotechnol Prog 35(1):e2743
Xu S, Gavin J, Jiang R, Chen H (2017) Bioreactor productivity and media cost comparison for different intensified cell culture processes. Biotechnol Prog 33(4):867–878
Matanguihan R et al (2001) Solution to the high dissolved CO2 problem in high-density perfusion culture of mammalian cells. In: Lindner-Olsson E, Chatzissavidou N, Lüllau E (eds) Animal cell technology: from target to market. ESACT proceedings, vol 1. Springer, Dordrecht
Acknowledgments
We would like to thank David Hacker for his input and support in writing this chapter and Eppendorf AG for providing images used in the creation of the figures.
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Schmid, A., Kreidl, E., Bertschinger, M., Vetsch, P. (2021). Benchtop Bioreactors in Mammalian Cell Culture: Overview and Guidelines. In: Turksen, K. (eds) Bioreactors in Stem Cell Biology. Methods in Molecular Biology, vol 2436. Humana, New York, NY. https://doi.org/10.1007/7651_2021_441
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DOI: https://doi.org/10.1007/7651_2021_441
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