title: Biologically active, magnICON®-expressed EPO-Fc from stably transformed Nicotiana benthamiana plants presenting tetra-antennary N-glycan structures.
abstract: In the past two decades plants have emerged as a valuable alternative for the production of pharmaceutical proteins. Since N-glycosylation influences functionality and stability of therapeutic proteins, the plant N-glycosylation pathway should be humanized. Here, we report the transient magnICON(®) expression of the erythropoietin fusion protein (EPO-Fc) in Nicotiana benthamiana plants that produce multi-antennary N-glycans without the plant-specific β1,2-xylose and α1,3-fucose residues in a stable manner (Nagels et al., 2011). The EPO-Fc fusion protein consists of EPO with a C-terminal-linked IgG-Fc domain and is used for pulmonary delivery of recombinant EPO to patients (Bitonti et al., 2004). Plant expressed EPO-Fc was quantified using a paramagnetic-particle chemiluminescent immunoassay and shown to be active in vitro via receptor binding experiments in HEK293T cells. Mass spectrometry-based N-glycan analysis confirmed the presence of multi-antennary N-glycans on plant-expressed EPO-Fc. The described research is the next step towards the development of a production platform for pharmaceutical proteins in plants.
authors.affiliation: Laboratory of Biochemistry and Glycobiology, Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium.
authors.forename: Bieke
authors.initials: B
authors.lastname: Nagels
authors.forename: Els J M
authors.initials: EJ
authors.lastname: Van Damme
authors.forename: Nico
authors.initials: N
authors.lastname: Callewaert
authors.forename: Lennart
authors.initials: L
authors.lastname: Zabeau
authors.forename: Jan
authors.initials: J
authors.lastname: Tavernier
authors.forename: Joris R
authors.initials: JR
authors.lastname: Delanghe
authors.forename: Annemie
authors.initials: A
authors.lastname: Boets
authors.forename: Alexandra
authors.initials: A
authors.lastname: Castilho
authors.forename: Koen
authors.initials: K
authors.lastname: Weterings
mesh_terms: D004921:Erythropoietin / Q000096:biosynthesis* / Q000235:genetics; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D011134:Polysaccharides / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D014026:Nicotiana / Q000502:physiology*; D014170:Transformation, Genetic / Q000235:genetics
chemical_list: D007128:Immunoglobulin Fragments; D011134:Polysaccharides; D004921:Erythropoietin