Plant Biotechnology, Faculty of Biology, University of Freiburg, Schaenzlestr. 1, 79104 Freiburg, Germany.
Sci Rep. 2013 Oct 22;3:3019. doi: 10.1038/srep03019.
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.
医药蛋白的重组生产不仅对个性化药物至关重要。虽然大多数生物制药目前都是在哺乳动物细胞培养中生产的，但植物制造的药物正在获得发展动力。植物中的翻译后修饰与人类相似，但现有的差异可能会影响产品的质量、安全性和疗效。高等真核生物中常见的修饰是脯氨酰-4-羟化酶（P4H）催化的脯氨酰羟化。靶蛋白上的 P4H 序列识别位点在人类和植物之间存在差异，导致在植物中生产的重组人蛋白发生非人类翻译后修饰。由此产生的羟脯氨酸显示出植物特异性 O-糖基化的锚点，对患者具有免疫原性。在这里，我们描述了一种负责植物（苔藓）生物反应器中重组人红细胞生成素（hEPO）的非人类脯氨酰羟化的植物基因的鉴定。该基因的靶向缺失消除了 hEPO 的不期望的脯氨酰羟化，从而为通过苔藓生物反应器中的糖基工程实现人源化的植物制造药物铺平了道路。
