title: Biochemical Characterization of Human Anti-Hepatitis B Monoclonal Antibody Produced in the Microalgae Phaeodactylum tricornutum.
abstract: 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 biopharmaceuticals. 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 microheterogeneity. 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.
authors.affiliation: Laboratoire Glycobiologie et Matrice Extracellulaire végétale Equipe d'Accueil 4358, Faculté des sciences et techniques, Université de Rouen, Normandie Université, Institut de Recherche et d'Innovation Biomédicale, Végétale Agronomie Sol Innovation, Mont-Saint-Aignan, France.
authors.forename: Gaëtan
authors.initials: G
authors.lastname: Vanier
authors.affiliation: LOEWE Center for Synthetic Microbiology, Philipps-Universität Marburg, Marburg, Germany.
authors.forename: Franziska
authors.initials: F
authors.lastname: Hempel
authors.affiliation: PISSARO Proteomic Platform, Normandie Université, Institut de Recherche et d'Innovation Biomédicale, Mont-Saint-Aignan, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Chan
authors.affiliation: Agilent Technologies, Waldbronn, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Rodamer
authors.affiliation: PISSARO Proteomic Platform, Normandie Université, Institut de Recherche et d'Innovation Biomédicale, Mont-Saint-Aignan, France.
authors.forename: David
authors.initials: D
authors.lastname: Vaudry
authors.affiliation: LOEWE Center for Synthetic Microbiology, Philipps-Universität Marburg, Marburg, Germany.
authors.forename: Uwe G
authors.initials: UG
authors.lastname: Maier
authors.affiliation: Laboratoire Glycobiologie et Matrice Extracellulaire végétale Equipe d'Accueil 4358, Faculté des sciences et techniques, Université de Rouen, Normandie Université, Institut de Recherche et d'Innovation Biomédicale, Végétale Agronomie Sol Innovation, Mont-Saint-Aignan, France.
authors.forename: Patrice
authors.initials: P
authors.lastname: Lerouge
authors.affiliation: Laboratoire Glycobiologie et Matrice Extracellulaire végétale Equipe d'Accueil 4358, Faculté des sciences et techniques, Université de Rouen, Normandie Université, Institut de Recherche et d'Innovation Biomédicale, Végétale Agronomie Sol Innovation, Mont-Saint-Aignan, France; Institut Universitaire de France, Paris, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Bardor
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods*; D017377:Diatoms / Q000378:metabolism*; D004355:Drug Stability; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006031:Glycosylation; D006510:Hepatitis B Antibodies / Q000096:biosynthesis*; D006514:Hepatitis B Surface Antigens / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis*; D013058:Mass Spectrometry; D058086:Microalgae / Q000378:metabolism*; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals; D021381:Protein Transport; D059748:Proteolysis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis
chemical_list: D000911:Antibodies, Monoclonal; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D007074:Immunoglobulin G; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D017351:N-Acetylglucosaminyltransferases; C024850:alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I