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Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-d-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes

Paper ID Volume ID Publish Year Pages File Format Full-Text
20293 43167 2014 7 PDF Available
Title
Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-d-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes
Abstract

Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Lea) epitope, Galβ(1–3)[Fucα(1–4)]GlcNAc. Because it is likely that these sugar residues and glycan structures are immunogenic, many attempts have been made to delete them. Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Lea epitopes by repressing the GDP-d-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-l-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. J., 9, 264–281, 2011). In the present study, we generated a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant). By crossing ΔGMD and ΔXylT plants, we successfully generated plants in which plant-specific sugar residues were repressed (ΔGMDΔXylT plants). The proportion of N-glycans with deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants increased by 82.41%. Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. Simultaneous repression of the GMD and XylT genes in N. benthamiana is thus very useful for deleting plant-specific sugar residues.

Keywords
Gene silencing; RNAi; Plant N-glycosylation; Recombinant proteins; Plant-specific sugar residues
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Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-d-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 118, Issue 4, October 2014, Pages 448–454
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us