中国仓鼠卵巢细胞
内质网
细胞培养
细胞生物学
重组DNA
单克隆抗体
效价
异源的
分子生物学
化学
抗体
基因
转染
生物化学
生物
免疫学
遗传学
作者
Katharine Cain,Shirley J. Peters,Hanna Hailu,Bernie Sweeney,Paul E. Stephens,James T. Heads,Kaushik Sarkar,Andy Ventom,C. Steven Page,Alan J. Dickson
摘要
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 ∼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-Lα) 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.
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