Biologically active, magnICON^?-expressed EPO-Fc from stably transformed Nicotiana benthamiana plants presenting tetra-antennary N-glycan structures

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• 作者：Bieke Nagels ; Els J.M. Van Damme ; Nico Callewaert ; Lennart Zabeau ; Jan Tavernier ; Joris R. Delanghe ; Annemie Boets ; Alex ; ra Castilho ; Koen Weterings
• 关键词：rhEPO ; recombinant human EPO ; GnT ; N-acetylglucosaminyltransferase ; XylT ; ¦Â1 ; 2-xylosyltransferase ; FucT ; ¦Á1 ; 3-fucosyltransferase ; EPOR ; EPO receptor ; LR ; leptin receptor ; LC&ndash ; ESI MS ; liquid chromatography&ndash ; electrospray ionization mass spectrom
• 刊名：Journal of Biotechnology
• 出版年：2012
• 出版时间：31 August, 2012
• 年：2012
• 卷：160
• 期：3-4
• 页码：242-250
• 全文大小：1117 K

文摘

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 ¦Â1,2-xylose and ¦Á1,3-fucose residues in a stable manner (). 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 (). 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.