Enhanced production of monomeric interferon-ß by CHO cells through the control of culture conditions
Enhanced production of monomeric interferon-ß by CHO cells through the control of culture conditions
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DOI：
10.1021/bp049807b
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发表时间：
2005-01-01
影响因子：
2.9
通讯作者：
Butler, M
中科院分区：
文献类型：
--
作者：
Rodriguez, J;Spearman, M;Butler, M
转染细胞系的重组蛋白表达的增强对于开发有效的大规模生物过程是必不可少的。研究了各种培养基添加剂和温度条件的影响，试图优化产人β-干扰素（Hu-beta-IFN）的中国仓鼠卵巢（CHO）细胞系的蛋白质产量、稳定性和蛋白质糖基化。我们观察到糖蛋白的ELISA反应在分批培养的后期阶段降低，这归因于分子聚集。在各种培养系统和条件下，将细胞置于各种浓度的甘油、二甲基亚砜（DMSO）和丁酸钠（NaBu）中。NaBu和DMSO的加入导致更高的比生产率，但降低了生长速率，导致产生的干扰素的净减少。甘油似乎稳定分泌的β-IFN，导致聚集减少，尽管细胞生长速率降低。分离的β-IFN的糖基化分析显示，在分批培养中唾液酸化的时间依赖性降低，甘油的存在改善了唾液酸化。低温条件（30摄氏度）对生产率的影响最大，β-干扰素滴度显着增加，分子聚集程度降低。
The enhancement of recombinant protein expression of a transfected cell line is essential for the development of an efficient large-scale bioprocess. The effect of various media additives and temperature conditions were studied in an attempt to optimize protein production, stability, and protein glycosylation from a Chinese hamster ovary (CHO) cell line producing human beta-interferon (Hu-beta-IFN). We observed a decrease in the ELISA response of the glycoprotein in the later stages of batch cultures, which was attributed to molecular aggregation. Cells were subjected to various concentrations of glycerol, dimethyl sulfoxide (DMSO), and sodium butyrate (NaBu) in a variety of culture systems and conditions. The addition of both NaBu and DMSO resulted in higher specific productivities but reduced growth rates that resulted in a net reduction of interferon produced. Glycerol appeared to stabilize the secreted beta-IFN, resulting in reduced aggregation, despite a decrease in cell growth rate. Glycosylation analysis of isolated beta-IFN showed a time-dependent decrease in sialylation in batch culture that was ameliorated by the presence of glycerol. Low-temperature conditions (30degreesC) had the greatest effect on productivity with a significant increase in beta-IFN titer as well as a reduction in the degree of molecular aggregation.