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Gewählte Publikation:

Strasser, R; Stadlmann, J; Schähs, M; Stiegler, G; Quendler, H; Mach, L; Glössl, J; Weterings, K; Pabst, M; Steinkellner, H.
(2008): Generation of glyco-engineered Nicotiana benthamiana for the production of monoclonal antibodies with a homogeneous human-like N-glycan structure.
Plant Biotechnol J. 2008; 6(4):392-402 FullText FullText_BOKU

Abstract:
A common argument against using plants as a production system for therapeutic proteins is their inability to perform authentic human N-glycosylation (i.e. the presence of beta 1,2-xylosylation and core alpha 1,3-fucosylation). In this study, RNA interference (RNAi) technology was used to obtain a targeted down-regulation of the endogenous beta 1,2-xylosyltransferase (XyIT) and alpha 1,3-fucosyltransferase (FucT) genes in Nicotiana benthamiana, a tobacco-related plant species widely used for recombinant protein expression. Three glyco-engineered lines with significantly reduced xylosylated and/or core alpha 1,3-fucosylated glycan structures were generated. The human anti HIV monoclonal antibody 2G12 was transiently expressed in these glycosylation mutants as well as in wild-type plants. Four glycoforms of 2G12 differing in the presence/absence of xylose and core alpha 1,3-fucose residues in their N-glycans were produced. Notably, 2G12 produced in XyIT/FucT-RNAi plants was found to contain an almost homogeneous N-glycan species without detectable xylose and alpha 1,3-fucose residues. Plant-derived glycoforms were indistinguishable from Chinese hamster ovary (CHO)-derived 2G12 with respect to electrophoretic properties, and exhibited functional properties (i.e. antigen binding and HIV neutralization activity) at least equivalent to those of the CHO counterpart. The generated RNAi lines were stable, viable and did not show any obvious phenotype, thus providing a robust tool for the production of therapeutically relevant glycoproteins in plants with a humanized N-glycan structure.
Autor*innen der BOKU Wien:
Glößl Josef
Mach Lukas
Pabst Martin
Quendler Heribert
Stadlmann Johannes
Steinkellner Herta
Strasser Richard
BOKU Gendermonitor:

Find related publications in this database (using NML MeSH Indexing)
Antibodies, Monoclonal - genetics
Down-Regulation -
Genetic Engineering -
Glycosylation -
HIV Antibodies - metabolism
HIV Antigens - metabolism
Humans -
Immunoglobulin G - metabolism
Mutation -
Polysaccharides - genetics
Protein Binding -
RNA Interference -
Tobacco - genetics

Find related publications in this database (Keywords)
2G12 anti-HIV antibody
Nicotiana benthamiana
plant N-glycosylation
recombinant proteins
RNAi glyco-engineering


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