Proteomics in biomanufacturing control: Protein dynamics of CHO-K1 cells and conditioned media during apoptosis and necrosis
Proteomics in biomanufacturing control: Protein dynamics of CHO-K1 cells and conditioned media during apoptosis and necrosis
复制标题
DOI：
10.1002/bit.26563
复制
发表时间：
2018-06-01
影响因子：
3.8
通讯作者：
Bones, Jonathan
中科院分区：
文献类型：
--
作者：
Albrecht, Simone;Kaisermayer, Christian;Bones, Jonathan
在治疗性蛋白质的生物制造过程中，细胞活力对产品的数量和质量有着至关重要的影响。因此，需要更深入地了解细胞和条件介质蛋白质组在细胞应激和死亡时的变化，以改进生物过程控制。本文应用高pH/低pH反相数据独立的2D-LC-MSE发现蛋白质组学平台，分别在台式生物反应器中用星形孢子素暴露或曝气剪切诱导细胞死亡的CHO-K1细胞和条件介质蛋白质组进行研究。对与分子功能、生物过程和细胞成分相关的基因本体论术语进行功能分类，揭示了细胞死亡的独立性和特殊性。此外，使用Hi3方法的无标记定量导致了23个潜在的细胞活力标记蛋白质的综合候选名单，这些蛋白质具有最高的丰度和诱导细胞死亡时条件培养液中显著增加的蛋白质，包括与细胞应激反应、信号调节、细胞骨架组织、细胞分化、细胞相互作用以及代谢和蛋白水解酶相关的蛋白质，这些蛋白质是转化为靶向分析平台用于监测生物处理反应和加强过程控制的有趣候选。
Cell viability has a critical impact on product quantity and quality during the biomanufacturing of therapeutic proteins. An advanced understanding of changes in the cellular and conditioned media proteomes upon cell stress and death is therefore needed for improved bioprocess control. Here, a high pH/low pH reversed phase data independent 2D-LC-MSE discovery proteomics platform was applied to study the cellular and conditioned media proteomes of CHO-K1 apoptosis and necrosis models where cell death was induced by staurosporine exposure or aeration shear in a benchtop bioreactor, respectively. Functional classification of gene ontology terms related to molecular functions, biological processes, and cellular components revealed both cell death independent and specific features. In addition, label free quantitation using the Hi3 approach resulted in a comprehensive shortlist of 23 potential cell viability marker proteins with highest abundance and a significant increase in the conditioned media upon induction of cell death, including proteins related to cellular stress response, signal mediation, cytoskeletal organization, cell differentiation, cell interaction as well as metabolic and proteolytic enzymes which are interesting candidates for translating into targeted analysis platforms for monitoring bioprocessing response and increasing process control.