Oh Han Kyu, So Moon Kyoung, Yang Jihye, Yoon Ho Chul, Ahn Ji Soo, Lee Jong Min, Kim Ji Tai, Yoo Ji Uk, Byun Tae Ho
Life Science Research Team, R&D Center, Samsung Fine Chemicals Co., Ltd., Rm. 6113, College of Medicine, SKKU 300, Chunchun-Dong, Jangan-Gu, Suwon, Kyunggi-Do, Korea.
Biotechnol Prog. 2005 Jul-Aug;21(4):1154-64. doi: 10.1021/bp050057v.
Sodium butyrate (NaBu) is used as a productivity enhancer for the production of therapeutic recombinant proteins in Chinese hamster ovary (CHO) cells. However, NaBu is well-known for having a cytotoxic effect, thereby inducing apoptosis. As an endeavor to reduce this defect, we studied 11 antioxidants known for inhibiting apoptosis, according to a Plackett-Burman statistical design on CHO cells producing recombinant interferon-beta-1a (IFN-beta). None of the antioxidants that we tested were as effective as N-acetylcystein (NAC) from the point of view of maintaining long-term survival of CHO cells and increasing the production of IFN-beta. In 7.5-L perfusion bioreactor cultures, the addition of NaBu and NAC elongated the culture period to almost 200 h throughout production phase and increased the production yield by 2-fold compared to control cultures containing only NaBu. Glycosylation patterns of produced IFN-beta at each run were also compared in IEF analysis. IEF profiles of where NaBu and NAC were added showed to be more isoforms with a lower pI than those of the control run. The sialic acid content was also increased by 17.7% according to HPLC analysis. Taken together, the data obtained demonstrate that the addition of NAC has positive effects on the elongation of the culture period, improving the production and increasing the sialylation of IFN-beta in NaBu-treated CHO cells.
丁酸钠（NaBu）被用作提高中国仓鼠卵巢（CHO）细胞生产治疗性重组蛋白产量的增强剂。然而，NaBu因具有细胞毒性作用，从而诱导细胞凋亡而闻名。为了减少这一缺陷，我们根据对生产重组β-1a干扰素（IFN-β）的CHO细胞进行的Plackett-Burman统计设计，研究了11种已知可抑制细胞凋亡的抗氧化剂。从维持CHO细胞长期存活和增加IFN-β产量的角度来看，我们测试的抗氧化剂中没有一种能像N-乙酰半胱氨酸（NAC）那样有效。在7.5-L灌注生物反应器培养中，与仅含有NaBu的对照培养物相比，添加NaBu和NAC使整个生产阶段的培养期延长至近200小时，并使产量提高了2倍。每次运行时所产生的IFN-β的糖基化模式也在等电聚焦分析中进行了比较。添加NaBu和NAC的等电聚焦图谱显示，与对照运行相比，具有更多低pI的异构体。根据高效液相色谱分析，唾液酸含量也增加了17.7%。综上所述，所获得的数据表明，添加NAC对延长NaBu处理的CHO细胞的培养期、提高产量以及增加IFN-β的唾液酸化具有积极作用。
