Skip to main content
SpringerLink
Log in
Book cover

Recombinant Proteins from Plants pp 161–172Cite as

Plant Cell-Based Recombinant Antibody Manufacturing with a 200 L Orbitally Shaken Disposable Bioreactor

  • Protocol

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1385))

Abstract

Tobacco BY-2 cells are an attractive platform for the manufacture of a variety of biopharmaceutical proteins, including antibodies. Here, we describe the scaled-up cultivation of human IgG–secreting BY-2 cells in a 200 L orbitally shaken disposable bioreactor, resulting in cell growth and recombinant protein yields that are proportionately comparable with those obtained from cultivations in 500 mL shake flasks. Furthermore, we present an efficient downstream process for antibody recovery from the viscous spent culture medium using expanded bed adsorption (EBA) chromatography.

This is a preview of subscription content, log in via an institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Christou P (2013) From medicinal plants to medicines in plants: plant factories for the production of valuable pharmaceuticals. Curr Pharm Des 19:5469–5470

    Article  CAS  PubMed  Google Scholar 

  2. Doran PM (2013) Therapeutically important proteins from in vitro plant tissue culture systems. Curr Med Chem 20:1047–1055

    CAS  PubMed  Google Scholar 

  3. Fischer R, Schillberg S, Buyel JF et al (2013) Commercial aspects of pharmaceutical protein production in plants. Curr Pharm Des 19:5471–5477

    Article  CAS  PubMed  Google Scholar 

  4. Twyman RM, Stoger E, Schillberg S et al (2003) Molecular farming in plants: host systems and expression technology. Trends Biotechnol 21:570–578

    Article  CAS  PubMed  Google Scholar 

  5. Basaran P, Rodríguez-Cerezo E (2008) Plant molecular farming: opportunities and challenges. Crit Rev Biotechnol 28:153–172

    Article  PubMed  Google Scholar 

  6. Hofbauer A, Stoger E (2013) Subcellular accumulation and modification of pharmaceutical proteins in different plant tissues. Curr Pharm Des 19:5495–5502

    Article  CAS  PubMed  Google Scholar 

  7. Häkkinen ST, Raven N, Henquet M et al (2014) Molecular farming in tobacco hairy roots by triggering the secretion of a pharmaceutical antibody. Biotechnol Bioeng 11:336–346

    Article  Google Scholar 

  8. Rival S, Wisniewski J-P, Langlais A et al (2008) Spirodela (duckweed) as an alternative production system for pharmaceuticals: a case study, aprotinin. Transgenic Res 17:503–513

    Article  CAS  PubMed  Google Scholar 

  9. Woodard SL, Wilken LR, Barros GOF et al (2009) Evaluation of monoclonal antibody and phenolic extraction from transgenic Lemna for purification process development. Biotechnol Bioeng 104:562–571

    Article  CAS  PubMed  Google Scholar 

  10. Hellwig S, Drossard J, Twyman RM et al (2004) Plant cell cultures for the production of recombinant proteins. Nat Biotechnol 22:1415–1422

    Article  CAS  PubMed  Google Scholar 

  11. Walmsley AM, Doran PM (2012) Foreign protein production using plant cells: opportunities and challenges. Biotechnol Adv 30:385–386

    Article  PubMed  Google Scholar 

  12. Nagata T, Nemoto Y (1992) Tobacco BY-2 cell line as the “HeLa” cell in the cell biology of higher plants. Int Rev Cytol 132:1–30

    Article  CAS  Google Scholar 

  13. Schillberg S, Raven N, Fischer R et al (2013) Molecular farming of pharmaceutical proteins using plant suspension cell and tissue cultures. Curr Pharm Des 19:5531–5542

    Article  CAS  PubMed  Google Scholar 

  14. Fischer R, Schillberg S, Hellwig S et al (2012) GMP issues for recombinant plant-derived pharmaceutical proteins. Biotechnol Adv 30:434–439

    Article  CAS  PubMed  Google Scholar 

  15. Eibl R, Kaiser S, Lombriser R et al (2010) Disposable bioreactors: the current state-of-the-art and recommended applications in biotechnology. Appl Microbiol Biotechnol 86:41–49

    Article  CAS  PubMed  Google Scholar 

  16. Eibl R, Eibl D (2006) Design and use of the wave bioreactor for plant cell culture. In: Gupta SD, Ibaraki Y (eds) Plant tissue culture engineering. Springer, Dordrecht, Netherlands, pp 203–227

    Google Scholar 

  17. Eibl R, Eibl D (2009) Design of bioreactors suitable for plant cell and tissue cultures. Phytochem Rev 7:593–598

    Article  Google Scholar 

  18. Georgiev MI, Eibl R, Zhong J-J (2013) Hosting the plant cells in vitro: recent trends in bioreactors. Appl Microbiol Biotechnol 97:3787–3800

    Article  CAS  PubMed  Google Scholar 

  19. Zhang X, Stettler M, De Sanctis D et al (2010) Use of orbital shaken disposable bioreactors for mammalian cell cultures from the milliliter-scale to the 1,000-liter scale. Adv Biochem Eng Biotechnol 115:33–53

    Google Scholar 

  20. Fairbanks G, Steck TL, Wallach DF (1971) Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10:2606–2617

    Article  CAS  PubMed  Google Scholar 

  21. Kirchhoff J, Raven N, Boes A et al (2012) Monoclonal tobacco cell lines with enhanced recombinant protein yields can be generated from heterogeneous cell suspension cultures by flow sorting. Plant Biotechnol J 10:936–944

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgement

This work was supported by the European Union Seventh Framework Program under grant agreement no. 227420 (CoMoFarm).

Author information

Authors and Affiliations

  1. Fraunhofer IME, Forckenbeckstraße 6, 52074, Aachen, Germany

    Nicole Raven, Stefan Schillberg & Stefan Rasche

Authors
  1. Nicole Raven
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Stefan Schillberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Stefan Rasche
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Stefan Schillberg .

Editor information

Editors and Affiliations

  1. Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, Ontario, Canada

    Jacqueline MacDonald

  2. Department of Biology, Western University, London, Ontario, Canada

    Igor Kolotilin

  3. London, Ontario, Canada

    Rima Menassa

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Raven, N., Schillberg, S., Rasche, S. (2016). Plant Cell-Based Recombinant Antibody Manufacturing with a 200 L Orbitally Shaken Disposable Bioreactor. In: MacDonald, J., Kolotilin, I., Menassa, R. (eds) Recombinant Proteins from Plants. Methods in Molecular Biology, vol 1385. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3289-4_12

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-3289-4_12

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3288-7

  • Online ISBN: 978-1-4939-3289-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation