The combined effect of sodium butyrate and low culture temperature on the production, sialylation, and biological activity of an antibody produced in CHO cells
The combined effect of sodium butyrate and low culture temperature on the production, sialylation, and biological activity of an antibody produced in CHO cells
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DOI：
10.1007/s12257-011-0069-8
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发表时间：
2011-12-01
影响因子：
3.2
通讯作者：
Tan, Wen-Song
中科院分区：
文献类型：
--
作者：
Chen, Fei;Kou, Tianci;Tan, Wen-Song
在30℃和37℃条件下培养含有0类似于5 mM丁酸钠（NaBu）的细胞，研究NaBu和低温对CHO细胞抗体产生数量和质量的联合影响。尽管在30℃和37℃下，NaBu的加入均以剂量依赖的方式降低细胞凋亡的活力，但降低培养温度可延缓NaBu诱导的显著凋亡的发生。在30A℃条件下，含2mm NaBu的培养物的特异性抗体产率（q (p)）最高，为23.26 pg/cell/day；然而，在30A℃下，含有1 mM NaBu的培养物中，抗体浓度最高，为167.84 mg/L，因为进一步添加NaBu对细胞生长的有害影响抵消了其对q的有益影响(p)。此外，还对总唾液酸含量和Nglycolylneuraminic acid （Neu5Gc）水平的蛋白质品质进行了评价。抗体的总唾液酸含量没有明显变化，但用NaBu处理或（和）低温培养确实使Neu5Gc水平降低了5%，约为10%。同时检测抗体的生物活性，未见明显变化。综上所述，在不影响唾液酸含量和生物活性的前提下，同时应用NaBu和低温培养是延长培养期和提高最终抗体浓度的有效方法。
Cell cultures containing 0 similar to 5 mM sodium butyrate (NaBu) and grown at 30 and 37A degrees C were conducted to investigate the combined effect of NaBu and low temperature on the quantity and quality of an antibody production in CHO cells. Although NaBu addition decreased cell viability by apoptosis in a dose-dependent manner at both 30 and 37A degrees C, the onset of significant apoptosis induced by NaBu was delayed by lowering culture temperature. The highest specific antibody productivity (q (p)) of 23.26 pg/cell/day was obtained in the culture containing 2 mM NaBu at 30A degrees C; however, the highest antibody concentration of 167.84 mg/L was achieved in the culture containing 1 mM NaBu at 30A degrees C, as the detrimental effect of further NaBu addition on cell growth compromised its beneficial effect on q (p). Moreover, protein quality with respect to the total sialic acid content and Nglycolylneuraminic acid (Neu5Gc) level was evaluated. There were no apparent changes regarding the total sialic acid content of the antibody, but manipulation of cultures with NaBu treatment or (and) low culture temperature did decrease Neu5Gc levels by 5 similar to 10%. Biological activity of the antibody was also assessed, and no obvious changes were observed. Collectively, the simultaneous application of NaBu and low culture temperature was an effective way to extend culture period and enhance final antibody concentration, without compromising the sialic acid content or biological activity.