Department of Biotechnology, Institute of Animal Cell Technology and Systems Biology (IACTSB), BOKU University, Muthgasse 18, 1190, Vienna, Austria.
Department of Biotechnology, Institute of Computational Biology (ICB), BOKU University, Muthgasse 18, 1190, Vienna, Austria.
Appl Microbiol Biotechnol. 2024 Jun 19;108(1):381. doi: 10.1007/s00253-024-13223-1.
Chinese hamster ovary (CHO) cells are popular in the pharmaceutical industry for their ability to produce high concentrations of antibodies and their resemblance to human cells in terms of protein glycosylation patterns. Current data indicate the relevance of CHO cells in the biopharmaceutical industry, with a high number of product commendations and a significant market share for monoclonal antibodies. To enhance the production capabilities of CHO cells, a deep understanding of their cellular and molecular composition is crucial. Genome sequencing and proteomic analysis have provided valuable insights into the impact of the bioprocessing conditions, productivity, and product quality. In our investigation, we conducted a comparative analysis of proteomic profiles in high and low monoclonal antibody-producing cell lines and studied the impact of tunicamycin (TM)-induced endoplasmic reticulum (ER) stress. We examined the expression levels of different proteins including unfolded protein response (UPR) target genes by using label-free quantification techniques for protein abundance. Our results show the upregulation of proteins associated with protein folding mechanisms in low producer vs. high producer cell line suggesting a form of ER stress related to specific protein production. Further, Hspa9 and Dnaja3 are notable candidates activated by the mitochondria UPR and play important roles in protein folding processes in mitochondria. We identified significant upregulation of Nedd8 and Lgmn proteins in similar levels which may contribute to UPR stress. Interestingly, the downregulation of Hspa5/Bip and Pdia4 in response to tunicamycin treatment suggests a low-level UPR activation. KEY POINTS: • Proteome profiling of recombinant CHO cells under mild TM treatment. • Identified protein clusters are associated with the unfolded protein response (UPR). • The compared cell lines revealed noticeable disparities in protein expression levels.
中国仓鼠卵巢（CHO）细胞因其能够产生高浓度的抗体以及在蛋白质糖基化模式方面与人细胞相似而在制药行业中广受欢迎。目前的数据表明 CHO 细胞在生物制药行业中的相关性，其产品推荐数量众多，单克隆抗体的市场份额也很大。为了提高 CHO 细胞的生产能力，深入了解其细胞和分子组成至关重要。基因组测序和蛋白质组学分析为了解生物加工条件、生产力和产品质量的影响提供了有价值的见解。在我们的研究中，我们对高产和低产单克隆抗体的 CHO 细胞系的蛋白质组图谱进行了比较分析，并研究了衣霉素（TM）诱导的内质网（ER）应激的影响。我们使用无标记定量技术研究了不同蛋白质的表达水平，包括未折叠蛋白反应（UPR）靶基因。我们的结果表明，与高产细胞系相比，低产细胞系中与蛋白质折叠机制相关的蛋白质上调，表明与特定蛋白质生产相关的某种形式的 ER 应激。此外，Hspa9 和 Dnaja3 是被线粒体 UPR 激活的重要候选蛋白，在蛋白质折叠过程中发挥重要作用。我们发现 Nedd8 和 Lgmn 蛋白的显著上调，其水平可能与 UPR 应激有关。有趣的是，Hspa5/Bip 和 Pdia4 的下调表明 UPR 激活程度较低。关键点：• 温和 TM 处理下重组 CHO 细胞的蛋白质组学分析。• 鉴定的蛋白质簇与未折叠蛋白反应（UPR）相关。• 比较细胞系显示蛋白质表达水平存在明显差异。
