title: A gene responsible for prolyl-hydroxylation of moss-produced recombinant human erythropoietin.
abstract: Recombinant production of pharmaceutical proteins is crucial, not only for personalized medicine. While most biopharmaceuticals are currently produced in mammalian cell culture, plant-made pharmaceuticals gain momentum. Post-translational modifications in plants are similar to those in humans, however, existing differences may affect quality, safety and efficacy of the products. A frequent modification in higher eukaryotes is prolyl-4-hydroxylase (P4H)-catalysed prolyl-hydroxylation. P4H sequence recognition sites on target proteins differ between humans and plants leading to non-human posttranslational modifications of recombinant human proteins produced in plants. The resulting hydroxyprolines display the anchor for plant-specific O-glycosylation, which bears immunogenic potential for patients. Here we describe the identification of a plant gene responsible for non-human prolyl-hydroxylation of human erythropoietin (hEPO) recombinantly produced in plant (moss) bioreactors. Targeted ablation of this gene abolished undesired prolyl-hydroxylation of hEPO and thus paves the way for plant-made pharmaceuticals humanized via glyco-engineering in moss bioreactors.
authors.affiliation: Plant Biotechnology, Faculty of Biology, University of Freiburg, Schaenzlestr. 1, 79104 Freiburg, Germany.
authors.forename: Juliana
authors.initials: J
authors.lastname: Parsons
authors.forename: Friedrich
authors.initials: F
authors.lastname: Altmann
authors.forename: Manuela
authors.initials: M
authors.lastname: Graf
authors.forename: Johannes
authors.initials: J
authors.lastname: Stadlmann
authors.forename: Ralf
authors.initials: R
authors.lastname: Reski
authors.forename: Eva L
authors.initials: EL
authors.lastname: Decker
mesh_terms: D019068:Bryopsida / Q000235:genetics* / Q000378:metabolism*; D000068817:Epoetin Alfa; D004921:Erythropoietin / Q000737:chemistry / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D006900:Hydroxylation; D042541:Intracellular Space / Q000378:metabolism; D013058:Mass Spectrometry; D011393:Procollagen-Proline Dioxygenase / Q000235:genetics* / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D004921:Erythropoietin; D000068817:Epoetin Alfa; D011393:Procollagen-Proline Dioxygenase