Johns Hopkins University , Baltimore, Maryland 21218, United States.
Brandeis University , Waltham, Massachusetts 02453, United States.
J Proteome Res. 2017 Oct 6;16(10):3672-3687. doi: 10.1021/acs.jproteome.7b00382. Epub 2017 Sep 14.
Chinese hamster ovary cells represent the dominant host for therapeutic recombinant protein production. However, few large-scale data sets have been generated to characterize this host organism and derived CHO cell lines at the proteomics level. Consequently, an extensive label-free quantitative proteomics analysis of two cell lines (CHO-S and CHO DG44) and two Chinese hamster tissues (liver and ovary) was used to identify a total of 11 801 unique proteins containing at least two unique peptides. 9359 unique proteins were identified specifically in the cell lines, representing a 56% increase over previous work. Additionally, 6663 unique proteins were identified across liver and ovary tissues, providing the first Chinese hamster tissue proteome. Protein expression was more conserved within cell lines during both growth phases than across cell lines, suggesting large genetic differences across cell lines. Overall, both gene ontology and KEGG pathway analysis revealed enrichment of cell-cycle activity in cells. In contrast, upregulated molecular functions in tissue include glycosylation and lipid transporter activity. Furthermore, cellular components including Golgi apparatus are upregulated in both tissues. In conclusion, this large-scale proteomics analysis enables us to delineate specific changes between tissues and cells derived from these tissues, which can help explain specific tissue function and the adaptations cells incur for applications in biopharmaceutical productions.
中国仓鼠卵巢细胞是治疗性重组蛋白生产的主要宿主。然而，很少有大规模数据集被生成来从蛋白质组学水平上对该宿主生物和衍生的 CHO 细胞系进行表征。因此，我们对两种细胞系（CHO-S 和 CHO DG44）和两种中国仓鼠组织（肝和卵巢）进行了广泛的无标记定量蛋白质组学分析，共鉴定出 11801 种具有至少两个独特肽段的独特蛋白质。9359 种独特蛋白质特异性地在细胞系中被鉴定出来，比以前的工作增加了 56%。此外，在肝和卵巢组织中鉴定出 6663 种独特蛋白质，提供了第一个中国仓鼠组织蛋白质组。在生长阶段，细胞系内的蛋白质表达比细胞系之间更保守，这表明细胞系之间存在较大的遗传差异。总体而言，基因本体论和 KEGG 途径分析都表明细胞周期活动在细胞中富集。相比之下，组织中上调的分子功能包括糖基化和脂质转运体活性。此外，高尔基体等细胞成分在两种组织中均上调。总之，这项大规模蛋白质组学分析使我们能够描绘出这些组织衍生的组织和细胞之间的具体变化，这有助于解释特定的组织功能以及细胞为生物制药生产而产生的适应。
