Institute for Chemical and Bio-Engineering (ICB), Swiss Federal Institute of Technology-ETH Zurich, Wolfgang-Pauli-Strasse 10,ETH Hoenggerberg, HCI F115, CH-8093 Zurich, Switzerland.
Metab Eng. 2006 May;8(3):264-72. doi: 10.1016/j.ymben.2006.01.006.
A variety of successful transcription and translation engineering strategies implemented during the past decade have driven the specific productivity of mammalian cells to an apparent limit. Restricted post-translation competence has since been considered the major bottleneck preventing mammalian cells from fully exploiting their physiologic production capacity in a biopharmaceutical manufacturing scenario. Through ectopic expression of the human transcription factor Xbp1 (X-box-binding-protein 1), evolved to manage plasma cell differentiation and coordinate the unfolded protein response, we have specifically expanded the endoplasmic reticulum and the Golgi of transgenic Chinese hamster ovary (CHO-K1)-derived cell lines with a resulting increase in overall production capacity. Xbp-1-based engineering of secretory bottlenecks was compatible with a variety of different promoter–product gene configurations suggesting that Xbp-1 induces generic production increases in CHO-K1 cell derivatives. Secretion engineering, illustrated here by Xbp1-based reprogramming of the post-translational processing machinery, provides a first insight into mastering a major system bottleneck which impacts biopharmaceutical manufacturing of secreted protein therapeutics.
在过去十年中，多种成功的转录和翻译工程策略已经将哺乳动物细胞的特定生产力推到了一个明显的极限。此后，翻译后能力的限制被认为是阻止哺乳动物细胞在生物制药生产环境中充分利用其生理生产能力的主要瓶颈。通过异位表达人类转录因子 Xbp1（X 盒结合蛋白 1），该因子进化为管理浆细胞分化并协调未折叠蛋白反应，我们专门扩展了内质网和转基因中国仓鼠卵巢（CHO-K1）衍生细胞系的高尔基体，从而提高了整体生产能力。基于 Xbp1 的分泌瓶颈工程与各种不同的启动子-产物基因构型兼容，表明 Xbp-1 诱导 CHO-K1 细胞衍生物的通用生产增加。分泌工程，如图所示，通过 Xbp1 对翻译后加工机制的重新编程，首次深入了解如何掌握影响分泌蛋白治疗剂生物制药生产的主要系统瓶颈。
