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Transient Glyco-Engineering of N. benthamiana Aiming at the Synthesis of Multi-antennary Sialylated Proteins

  • Andreas Loos
  • Alexandra Castilho
Part of the Methods in Molecular Biology book series (MIMB, volume 1321)

Abstract

Recombinantly produced therapeutic proteins bring benefits to patients and production companies alike. However, due to high production costs the potential of this technology cannot be fully tapped and therefor safe, scalable, and economic alternatives to the standard mammalian cell culture-based manufacturing systems are needed. Plant-based expression systems with their current technological advances constitute such an alternative. Many recombinant biopharmaceuticals are glycoproteins and their structural properties and pharmacokinetics are strongly influenced by their glycosylation profile. Differences in glycosylation between plants and mammals can for this reason result in different therapeutic efficacies. In particular, low levels of sialylation may lead to a short serum half-life of therapeutic proteins and nonhuman types of glycosylation can induce degradation and immunogenic responses. Controlling glycosylation of plant-derived therapeutics is therefore fundamental to enhance their efficacy and eliminate possible adverse effects caused by non-authentic glycosylation. Here we describe methods to transiently express high levels of recombinant proteins in Nicotiana benthamiana and simultaneously modulate their glycosylation pattern towards the synthesis of highly sialylated humanlike structures.

Key words

N-glycosylation Glyco-engineering In planta sialylation Transient expression N. benthamiana Molecular farming 

Notes

Acknowledgments

This work was supported by a grant from the Austrian Research Promotion Agency (FFG) Laura Bassi Centre of Expertise PlantBioP (Number 822757), and the Austrian Science Fund (FWF; Grant Number L575-B13).

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Andreas Loos
    • 1
  • Alexandra Castilho
    • 1
  1. 1.Department of Applied Genetics and Cell BiologyUniversity of Natural Resources and Life SciencesViennaAustria

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