Meleady Paula, Henry Michael, Gammell Patrick, Doolan Padraig, Sinacore Martin, Melville Mark, Francullo Linda, Leonard Mark, Charlebois Timothy, Clynes Martin
National Institute for Cellular Biotechnology, Dublin City University, Dublin, Ireland.
Proteomics. 2008 Jul;8(13):2611-24. doi: 10.1002/pmic.200700854.
Chinese hamster ovary (CHO) cells are widely used for the production of recombinant protein biopharmaceuticals. The purpose of this study was to investigate differences in the proteome of CHO DUKX cells expressing recombinant human bone morphogenetic protein-2 (rhBMP-2) (G5 cells) compared to cells also expressing soluble exogenous paired basic amino acid cleaving enzyme soluble paired basic amino acid cleaving enzyme (PACEsol) (3C9 cells), which has been previously found to improve the post-translational processing of the mature rhBMP-2 dimer. PACEsol co-expression was also associated with a significant increase (almost four-fold) in cellular productivity of rhBMP-2 protein. Differential proteomic expression profiling using 2-D DIGE and MALDI-TOF MS was performed to compare 3C9 and G5 cells, and revealed a list of 60 proteins that showed differential expression (up/downregulated), with a variety of different cellular functions. A substantial number of these altered proteins were found to have chaperone activity, involved with protein folding, assembly and secretion, as well as a number of proteins involved in protein translation. These results support the use of proteomic profiling as a valuable tool towards understanding the biology of bioprocess cultures.
中国仓鼠卵巢（CHO）细胞被广泛用于重组蛋白生物制药的生产。本研究的目的是调查表达重组人骨形态发生蛋白-2（rhBMP-2）的CHO DUKX细胞（G5细胞）与同样表达可溶性外源成对碱性氨基酸裂解酶（PACEsol）的细胞（3C9细胞）蛋白质组的差异，此前已发现PACEsol可改善成熟rhBMP-2二聚体的翻译后加工。PACEsol共表达还与rhBMP-2蛋白的细胞生产力显著提高（几乎四倍）相关。使用二维差异凝胶电泳（2-D DIGE）和基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）进行差异蛋白质组表达谱分析，以比较3C9和G5细胞，并揭示了60种显示差异表达（上调/下调）的蛋白质列表，这些蛋白质具有多种不同的细胞功能。发现大量这些改变的蛋白质具有伴侣活性，参与蛋白质折叠、组装和分泌，以及一些参与蛋白质翻译的蛋白质。这些结果支持将蛋白质组分析作为理解生物过程培养生物学的有价值工具。
