Subjects: Biology >> Bioengineering Subjects: Engineering and technical science >> Bioengineering submitted time 2018-12-11
Abstract： Carbon dioxide partial pressure (pCO2) is an important parameter in mammalian cell culture, which come from cell respiration, bicarbonate in culture media and CO2 added during culture for pH maintenance. Generally, pCO2 is not precisely controlled like pH in cell culture, but it is crucial to maintain pCO2 level within a reasonable range, usually ranging from 30 to 80 mmHg. Higher (> 80mmHg) or lower (< 30mmHg) pCO2 levels were reported to be detrimental to cell growth, metabolism, productivity and product quality. Higher pCO2 is always encountered during scale-up, especially for the high density perfusion culture. Therefore, pCO2 difference between small scale (< 15 L) and large scale (> 200L) needs to be considered. This review illustrates the effects of pCO2 on cell growth, metabolism, productivity and product quality. In addition, the approaches to control pCO2 during scale-up are also discussed. " "
Peer Review Status:
Awaiting Review
Subjects: Biology >> Bioengineering submitted time 2018-11-08 Cooperative journals: 《中国生物工程杂志》
Abstract：当前生物制药领域，由于成本压力、市场需求急剧波动以及生物仿制药的竞争日益激烈，现有的生物制造技术受到诸多挑战，生物技术公司越来越倾向于开发灵活、高效的创新型生产制造工艺。灌流培养作为当前哺乳动物细胞培养的重要工艺之一，不仅可以通过不断移出副产物和添加营养物来提供有利于细胞的稳定环境，以解决蛋白质量不稳定或者表达量偏低等问题，还可以通过提高单位体积产率来优化产能利用率并提高生产效率。本文就灌流培养用于哺乳动物细胞培养的研究进展做了系统介绍，并为其进一步开发与应用提供参考。
Subjects: Biology >> Bioengineering submitted time 2018-05-23 Cooperative journals: 《中国生物工程杂志》
Abstract：目的：筛选高表达单克隆细胞株，并通过优化培养基及流加物，最终达到提高目的蛋白产量及质量的目的。方法：通过有限稀释法对转染目的蛋白的CHO-S细胞进行单克隆化，应用双抗夹心ELISA方法对单克隆细胞株抗体表达量进行初步评估，最后根据筛选细胞株的活率、密度、产量及代谢情况，选择2-3株单克隆细胞进行培养条件优化，并对获得的发酵液进行纯化捕获，根据抗体蛋白表达量、糖型、等电点、纯度、酸碱峰分布等进行相应的评估分析，筛选出最优细胞株及最优培养方案。结果：经过单克隆化处理以及培养条件优化，蛋白的表达量由初始的不到500mg/L提升到2290mg/L ，且抗体蛋白纯度高达97.48%。抗体蛋白质量分析结果显示B1方案为本次实验最优培养方案。结论：通过细胞株筛选、培养基优化能显著提高抗体蛋白的产量及质量，同时对抗体蛋白糖型、等电点、纯度等均有一定程度的优化。因此工业生产中我们完全可以通过高表达克隆的筛选、培养工艺优化等对目的蛋白产量及质量进行一定程度的改善与提高，本次研究对后期实验研究及工业化方案开发都具有很好的指导意义。
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