Biochemical Characterization of Human Anti- Hepatitis B Monoclonal Antibody Produced in the Microalgae Phaeodactylum tricornutum

Gaëtan Vanier (1) , Franziska Hempel (2) , Philippe Chan (3) , Michael Rodamer (4) , David Vaudry (3) , Uwe Maier (2) , Patrice Lerouge (1) , Muriel Bardor (1)

1 Glyco-MEV - Laboratoire de Glycobiologie et Matrice Extracellulaire Végétale
2 Philipps Universität Marburg = Philipps University of Marburg
3 Plate-forme de Protéomique PISSARO
4 Agilent Technologies France

Monoclonal antibodies (mAbs) represent actually the major class of biopharmaceuticals. They are produced recombinantly using living cells as biofactories. Among the different expression systems currently available, microalgae represent an emerging alternative which displays several biotechnological advantages. Indeed, microalgae are classified as generally recognized as safe organisms and can be grown easily in bioreactors with high growth rates similarly to CHO cells. Moreover, microalgae exhibit a phototrophic lifestyle involving low production costs as protein expression is fueled by photosynthesis. However, questions remain to be solved before any industrial production of algae-made biopharma-ceuticals. Among them, protein heterogeneity as well as protein post-translational modifications need to be evaluated. Especially, N-glycosylation acquired by the secreted recombinant proteins is of major concern since most of the biopharmaceuticals including mAbs are N-glycosylated and it is well recognized that glycosylation represent one of their critical quality attribute. In this paper, we assess the quality of the first recombinant algae-made mAbs produced in the diatom, Phaeodactylum tricornutum. We are focusing on the characterization of their C-and N-terminal extremities, their signal peptide cleavage and their post-translational modifications including N-glycosylation macro-and microhetero-geneity. This study brings understanding on diatom cellular biology, especially secretion and intracellular trafficking of proteins. Overall, it reinforces the positioning of P. tricornutum as an emerging host for the production of biopharmaceuticals and prove that P. tricornutum is suitable for producing recombinant proteins bearing high mannose-type N-glycans.

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]

Gaëtan Vanier, Franziska Hempel, Philippe Chan, Michael Rodamer, David Vaudry, et al.. Biochemical Characterization of Human Anti- Hepatitis B Monoclonal Antibody Produced in the Microalgae Phaeodactylum tricornutum. PLoS ONE, 2015, 10 (10), pp.e0139282. ⟨10.1371/journal.pone.0139282⟩. ⟨hal-01841078⟩