An XBP-1 dependent bottle-neck in production of IgG subtype antibodies in chemically defined serum-free Chinese hamster ovary (CHO) fed-batch processes
An XBP-1 dependent bottle-neck in production of IgG subtype antibodies in chemically defined serum-free Chinese hamster ovary (CHO) fed-batch processes
复制标题
DOI：
10.1016/j.jbiotec.2008.03.008
复制
发表时间：
2008-06-01
影响因子：
4.1
通讯作者：
Kaufmann, Hitto
中科院分区：
文献类型：
--
作者：
Becker, Eric;Florin, Lore;Kaufmann, Hitto
治疗性抗体生产工艺的优化是制药生物技术领域的持续挑战。尽管可以证明载体设计和宿主细胞工程可以提高转录和翻译效率，从而产生高产细胞系，但尚不清楚引入调节蛋白质转运或影响翻译后修饰的转基因是否可以进一步改善这种工业过程。在这里，我们证明转录因子X-box结合蛋白-1（XBP-1）的异源表达可以导致中国仓鼠卵巢内质网（ER）含量和特异性治疗抗体生产力的增加接种悬浮培养物中的 (CHO)-DG44 细胞。这种效应转化为补料分批形式的总体抗体滴度增加 40%，其中细胞在化学成分确定的无血清培养基中生长。发现来自模拟转染细胞和 XBP-1 转染细胞的 Protein-A 纯化抗体产品在糖基化模式和理化特征方面具有相当的质量。这些数据证明了 XBP-1 工程在改善哺乳动物细胞培养生产过程以产生大量具有所需质量的治疗性蛋白质产品方面的潜力。 (c) 2008 Elsevier B.V. 保留所有权利。
The optimization of production processes for therapeutic antibodies is a continuing challenge in pharmaceutical biotechnology. Although it could be demonstrated that vector design and host cell engineering can improve transcriptional and translational efficiency and thereby result in generation of high producer cell lines, it is not clear whether introduction of transgenes that regulate protein transport or affect post-translational modifications could further improve such industrial processes.Here, we show that heterologous expression of the transcription factor X-box binding protein-1 (XBP-1) can lead to an increase in endoplasmic reticulum (ER) content and specific therapeutic antibody productivity of Chinese hamster ovary (CHO)-DG44 cells in inoculum suspension cultures. This effect translates into 40% increased overall antibody titers in a fed-batch format where cells are grown in chemically defined serum-free media. Protein-A purified antibody products from mock-transfected cells and XBP-1 transfected cells were found to be of comparable quality with regard to glycosylation pattern and physicochemical characteristics. The data demonstrate the potential of XBP-1 engineering to improve mammalian cell culture production processes to yield high amounts of a therapeutic protein product of desired quality. (c) 2008 Elsevier B.V. All rights reserved.