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Brassica rapa hairy root based expression system leads to the production of highly homogenous and reproducible profiles of recombinant human alpha-L-iduronidase.
- Source :
-
Plant biotechnology journal [Plant Biotechnol J] 2019 Feb; Vol. 17 (2), pp. 505-516. Date of Electronic Publication: 2018 Aug 30. - Publication Year :
- 2019
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Abstract
- The Brassica rapa hairy root based expression platform, a turnip hairy root based expression system, is able to produce human complex glycoproteins such as the alpha-L-iduronidase (IDUA) with an activity similar to the one produced by Chinese Hamster Ovary (CHO) cells. In this article, a particular attention has been paid to the N- and O-glycosylation that characterize the alpha-L-iduronidase produced using this hairy root based system. This analysis showed that the recombinant protein is characterized by highly homogeneous post translational profiles enabling a strong batch to batch reproducibility. Indeed, on each of the 6 N-glycosylation sites of the IDUA, a single N-glycan composed of a core Man <subscript>3</subscript> GlcNAc <subscript>2</subscript> carrying one beta(1,2)-xylose and one alpha(1,3)-fucose epitope (M3XFGN2) was identified, highlighting the high homogeneity of the production system. Hydroxylation of proline residues and arabinosylation were identified during O-glycosylation analysis, still with a remarkable reproducibility. This platform is thus positioned as an effective and consistent expression system for the production of human complex therapeutic proteins.<br /> (© 2018 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.)
- Subjects :
- Animals
Brassica rapa genetics
CHO Cells
Cricetulus
Epitopes immunology
Fucose immunology
Glycosylation
Humans
Iduronidase chemistry
Iduronidase genetics
Mannose metabolism
Plant Roots enzymology
Plant Roots genetics
Plants, Genetically Modified
Polysaccharides metabolism
Recombinant Proteins
Reproducibility of Results
Transgenes
Xylose immunology
Brassica rapa enzymology
Iduronidase metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1467-7652
- Volume :
- 17
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Plant biotechnology journal
- Publication Type :
- Academic Journal
- Accession number :
- 30058762
- Full Text :
- https://doi.org/10.1111/pbi.12994