A CHO cell line engineered to express XBP1 and ERO1-L has increased levels of transient protein expression
A CHO cell line engineered to express XBP1 and ERO1-L has increased levels of transient protein expression
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DOI：
10.1002/btpr.1693
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发表时间：
2013-05-01
影响因子：
2.9
通讯作者：
Dickson, Alan
中科院分区：
文献类型：
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作者：
Cain, Katharine;Peters, Shirley;Dickson, Alan
瞬时基因表达（TGE）系统目前提供了用于产生多克量的重组异源蛋白的快速且可扩展（高达100 L）的方法。已在HEK 293细胞中证明了高达1 g/L的产物滴度，但中国仓鼠卵巢（CHO）细胞的报告产量较低，约为300 mg/L。我们报告的建立工程CHOS细胞系，这已被开发为TGE。该细胞系已被工程化以表达X-box结合蛋白（XBP-1 S）和内质网氧化还原酶（ERO 1-L），并被命名为CHOS-XE。与CHOS细胞相比，CHOS-XE细胞产生增加的抗体（MAb）产量（5.3- 6.2倍）。通过尺寸、电荷、聚集倾向、主要糖基化物质和热稳定性评估，产品质量未发生变化。为了进一步开发和测试该TGE系统，评估了五种商业培养基，其中一种显示出抗体产量的最大增加。在添加商业饲料的情况下，单克隆抗体滴度达到875 mg/L。(c)2013年美国化学工程师学会生物技术。程序：29：697-706，2013
Transient gene expression (TGE) systems currently provide rapid and scalable (up to 100 L) methods for generating multigram quantities of recombinant heterologous proteins. Product titers of up to 1 g/L have been demonstrated in HEK293 cells but reported yields from Chinese hamster ovary (CHO) cells are lower at approximate to 300 mg/L. We report on the establishment of an engineered CHOS cell line, which has been developed for TGE. This cell line has been engineered to express both X-box binding protein (XBP-1S) and endoplasmic reticulum oxidoreductase (ERO1-L) and has been named CHOS-XE. CHOS-XE cells produced increased antibody (MAb) yields (5.3- 6.2 fold) in comparison to CHOS cells. Product quality was unchanged as assessed by size, charge, propensity to aggregate, major glycosylation species, and thermal stability. To further develop and test this TGE system, five commercial media were assessed, and one was shown to offer the greatest increase in antibody yields. With the addition of a commercial feed, MAb titers reached 875 mg/L. (c) 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:697-706, 2013