Application of 13C flux analysis to identify high-productivity CHO metabolic phenotypes.
Application of 13C flux analysis to identify high-productivity CHO metabolic phenotypes.
复制标题
应用 13C 通量分析来识别高产率 CHO 代谢表型。
DOI：
10.1016/j.ymben.2017.01.008
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发表时间：
2017
影响因子：
8.4
通讯作者：
Jamey D. Young
中科院分区：
文献类型：
--
作者：
Neil Templeton;Kevin D. Smith;Allison G McAtee;H. Dorai;M. Betenbaugh;Steven E Lang;Jamey D. Young
工业生物过程对宿主细胞的能量代谢提出了很高的要求，以满足最大蛋白质表达的生物合成要求。因此，鉴定促进高表达的代谢表型是生物技术工业的主要目标。我们进行了一系列13 C通量分析研究，以检查在3 L补料分批生物反应器中生长的CHO细胞培养物的早期稳定期期间对IgG表达的代谢反应。我们检查了八个克隆表达四种不同的IgG，并与三个非表达宿主细胞对照。一些克隆通过表达Bcl-2Δ进行遗传操作以使其具有抗凋亡性，Bcl-2 Δ与增加的IgG产生和升高的葡萄糖代谢相关。通过分层聚类分析将非表达、IgG表达和Bcl-2Δ/IgG表达克隆的代谢表型完全分离。乳酸消耗和柠檬酸循环通量与特异性IgG生产率最密切相关。这些研究表明，氧化代谢增强是高产CHO细胞系的一个特征。
Industrial bioprocesses place high demands on the energy metabolism of host cells to meet biosynthetic requirements for maximal protein expression. Identifying metabolic phenotypes that promote high expression is therefore a major goal of the biotech industry. We conducted a series of13C flux analysis studies to examine the metabolic response to IgG expression during early stationary phase of CHO cell cultures grown in 3 L fed-batch bioreactors. We examined eight clones expressing four different IgGs and compared with three non-expressing host-cell controls. Some clones were genetically manipulated to be apoptosis-resistant by expressing Bcl-2Δ, which correlated with increased IgG production and elevated glucose metabolism. The metabolic phenotypes of the non-expressing, IgG-expressing, and Bcl-2Δ/IgG-expressing clones were fully segregated by hierarchical clustering analysis. Lactate consumption and citric acid cycle fluxes were most strongly associated with specific IgG productivity. These studies indicate that enhanced oxidative metabolism is a characteristic of high-producing CHO cell lines.
影响因子：
7.7
作者：
Young JD
通讯作者：
Young JD