Brassica rapa hairy root based expression system leads to the production of highly homogenous and reproducible profiles of recombinant human alpha-L-iduronidase

Florian Cardon (1) , Roser Pallisse (1) , Muriel Bardor (2, 3) , Aurore Caron (1) , Jessica Vanier (2) , Jean-Pierre Ele Ekouna (4) , Patrice P. Lerouge (2) , Michèle Boitel-Conti (4) , Marina Guillet (1)

1 Root Lines Technology
2 Glyco-MEV - Laboratoire de Glycobiologie et Matrice Extracellulaire Végétale
3 IUF - Institut Universitaire de France
4 BIOPI - Biologie des Plantes et Innovation - UR UPJV 3900

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 Man3GlcNAc2 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.

Keywords: post‐translational modification; glycosylation; alpha—L—iduronidase; hairy root; plant based expression system; recombinant; glycoprotein; alpha-L- iduronidase; post-translational modification

Domains: Chemical Sciences; Polymers; Life Sciences [q-bio]; Vegetal Biology; Plant breeding; Life Sciences [q-bio]; Cellular Biology; Cell Behavior [q-bio.CB]; Life Sciences [q-bio]; Biochemistry, Molecular Biology; Biochemistry [q-bio.BM]; Life Sciences [q-bio]; Biochemistry, Molecular Biology; Molecular biology; Life Sciences [q-bio]; Biochemistry, Molecular Biology; Genomics [q-bio.GN]; Life Sciences [q-bio]; Biochemistry, Molecular Biology; Molecular Networks [q-bio.MN]; Life Sciences [q-bio]; Vegetal Biology; Phytopathology and phytopharmacy; Life Sciences [q-bio]; Biotechnology; Life Sciences [q-bio]; Development Biology; Gametogenesis; Life Sciences [q-bio]; Cellular Biology; Subcellular Processes [q-bio.SC]

Florian Cardon, Roser Pallisse, Muriel Bardor, Aurore Caron, Jessica Vanier, et al.. Brassica rapa hairy root based expression system leads to the production of highly homogenous and reproducible profiles of recombinant human alpha-L-iduronidase. Plant Biotechnology Journal, In press, 17 (2), pp.505-516. ⟨10.1111/pbi.12994⟩. ⟨hal-01873252⟩