Author:
Korn Janin,Schäckermann Dorina,Kirmann Toni,Bertoglio Federico,Steinke Stephan,Heisig Janyn,Ruschig Maximilian,Rojas Gertrudis,Langreder Nora,Wenzel Esther Veronika,Roth Kristian Daniel Ralph,Becker Marlies,Meier Doris,van den Heuvel Joop,Hust Michael,Dübel Stefan,Schubert Maren
Abstract
AbstractAntibodies are essential tools for therapy and diagnostics. Yet, production remains expensive as it is mostly done in mammalian expression systems. As most therapeutic IgG require mammalian glycosylation to interact with the human immune system, other expression systems are rarely used for production. However, for neutralizing antibodies that are not required to activate the human immune system as well as antibodies used in diagnostics, a cheaper production system would be advantageous. In our study, we show cost-efficient, easy and high yield production of antibodies as well as various secreted antigens including Interleukins and SARS-CoV-2 related proteins in a baculovirus-free insect cell expression system. To improve yields, we optimized the expression vector, media and feeding strategies. In addition, we showed the feasibility of lyophilization of the insect cell produced antibodies. Furthermore, stability and activity of the antibodies was compared to antibodies produced by Expi293F cells revealing a lower aggregation of antibodies originating from High Five cell production. Finally, the newly established High Five expression system was compared to the Expi293F mammalian expression system in regard of yield and costs. Most interestingly, all tested proteins were producible in our High Five cell expression system what was not the case in the Expi293F system, hinting that the High Five cell system is especially suited to produce difficult-to-express target proteins.
Funder
Alexander von Humboldt-Stiftung
Projekt DEAL
Publisher
Springer Science and Business Media LLC
Reference39 articles.
1. Monoclonal Antibodies Market - Forecasts from 2018 to 2023. Resarch and Markets https://www.researchandmarkets.com/reports/4618236/monoclonal-antibodies-market-forecasts-from (2018).
2. Chadd, H. E. & Chamow, S. M. Therapeutic antibody expression technology. Curr. Opin. Biotechnol. 12, 188–194 (2001).
3. Kelley, B. Very large scale monoclonal antibody purification: the case for conventional unit operations. Biotechnol. Prog. 23, 995–1008 (2007).
4. Kelley, B. Industrialization of mAb production technology: the bioprocessing industry at a crossroads. mAbs 1, 443–452 (2009).
5. Vink, T., Oudshoorn-Dickmann, M., Roza, M., Reitsma, J.-J. & de Jong, R. N. A simple, robust and highly efficient transient expression system for producing antibodies. Methods 65, 5–10 (2014).
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