Metabolite profiling of CHO cells: Molecular reflections of bioprocessing effectiveness.
Metabolite profiling of CHO cells: Molecular reflections of bioprocessing effectiveness.
复制标题
DOI：
10.1002/biot.201400664
复制
发表时间：
2015-09
影响因子：
4.7
通讯作者：
C. Sellick;Alexandra S. Croxford;A. Maqsood;G. Stephens;H. Westerhoff;R. Goodacre;A. Dickson
中科院分区：
文献类型：
--
作者：
C. Sellick;Alexandra S. Croxford;A. Maqsood;G. Stephens;H. Westerhoff;R. Goodacre;A. Dickson
虽然培养基和补料的开发为CHO细胞中的重组蛋白生产提供了重大进展，但基本的理解是有限的。我们已经应用代谢物分析与建立健全的（GC-MS）分析，以确定分子位点，通过该分子位点，两个产量提高饲料提高重组抗体产量从一个模型GS-CHO细胞系。通过跨核心代谢途径的数据，这些数据报告了几个细胞隔室中的代谢，这些数据确定了与细胞存活率和细胞比生产率增加相关的关键代谢物和事件。特别重要的是，增加的过程效率与线粒体的功能活性有关，其中柠檬酸循环的中间体的使用/产生的量和时间过程，用于诸如脂质生物合成、前体产生和能量产生的用途，提供了关于生产力的细胞状态的直接指标。这些数据提供了特定细胞代谢指标和细胞过程效率之间的明确关联，从乳酸代谢平衡与其他氧化还原汇（甘油，山梨醇和苏糖醇）的相关性的先前指示延伸。这些信息及其解释从遗传或环境角度确定了工程细胞培养效率的目标，并更好地了解特定培养基组分对整个CHO细胞生物加工的重要性。
Whilst development of medium and feeds has provided major advances in recombinant protein production in CHO cells, the fundamental understanding is limited. We have applied metabolite profiling with established robust (GC-MS) analytics to define the molecular loci by which two yield-enhancing feeds improve recombinant antibody yields from a model GS-CHO cell line. With data across core metabolic pathways, that report on metabolism within several cellular compartments, these data identify key metabolites and events associated with increased cell survival and specific productivity of cells. Of particular importance, increased process efficiency was linked to the functional activity of the mitochondria, with the amount and time course of use/production of intermediates of the citric acid cycle, for uses such as lipid biosynthesis, precursor generation and energy production, providing direct indicators of cellular status with respect to productivity. The data provide clear association between specific cellular metabolic indicators and cell process efficiency, extending from prior indications of the relevance of lactate metabolic balance to other redox sinks (glycerol, sorbitol and threitol). The information, and its interpretation, identifies targets for engineering cell culture efficiency, either from genetic or environmental perspectives, and greater understanding of the significance of specific medium components towards overall CHO cell bioprocessing.