Abstract
Recombinant glycoproteins such as monoclonal antibodies have a major impact on modern healthcare systems, e.g., as the active pharmaceutical ingredients in anticancer drugs. A specific glycan profile is often necessary to achieve certain desirable activities, such as the effector functions of an antibody, receptor binding or a sufficient serum half-life. However, many expression systems produce glycan profiles that differ substantially from the preferred form (usually the form found in humans) or produce a diverse array of glycans with a range of in vivo activities, thus necessitating laborious and costly separation and purification processes. In contrast, protein glycosylation in plant cells is much more homogeneous than other systems, with only one or two dominant forms. Additionally, these glycan profiles tend to remain stable when the process and cultivation conditions are changed, making plant cells an ideal expression system to produce recombinant glycoproteins with uniform glycan profiles in a consistent manner. This chapter describes a protocol that uses fermentations using plant cell cultures to produce glycosylated proteins using two different types of bioreactors, a classical autoclavable STR 3-L and a wave reactor.
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Abbreviations
- cdO2 :
-
Controller output for the oxygen control loop
- dO2 :
-
Dissolved oxygen
- DW:
-
Dry weight
- FW:
-
Fresh weight
- OUR:
-
Oxygen uptake rate
- PCV:
-
Packed cell volume
- RT:
-
Room temperature
- STR:
-
Stirred tank reactor
- VVM:
-
Volume per volume and minutes
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Acknowledgments
The authors acknowledge Dr. Richard M Twyman for editorial assistance. This work was funded in part the Fraunhofer-Gesellschaft Internal Programs under Grant No. Attract 125-600164. The authors have no conflict of interest to declare.
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Holland, T., Buyel, J.F. (2018). Bioreactor-Based Production of Glycoproteins in Plant Cell Suspension Cultures. In: Picanço-Castro, V., Swiech, K. (eds) Recombinant Glycoprotein Production. Methods in Molecular Biology, vol 1674. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7312-5_11
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DOI: https://doi.org/10.1007/978-1-4939-7312-5_11
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