Strategies for selecting recombinant CHO cell lines for cGMP manufacturing: Improving the efficiency of cell line generation
Strategies for selecting recombinant CHO cell lines for cGMP manufacturing: Improving the efficiency of cell line generation
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选择用于 cGMP 生产的重组 CHO 细胞系的策略：提高细胞系生成的效率
DOI：
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发表时间：
2010
期刊：
影响因子：
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通讯作者：
A. Dickson
中科院分区：
文献类型：
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作者：
Alison J. Porter;A. Racher;R. Preziosi;A. Dickson
在细胞系生成过程中获得了具有广泛细胞表型的转染物。为了成功制造治疗性蛋白，需要一种方法从这种表型广泛的转染群体中鉴定出具有理想生长和生产力特征的细胞系。这一鉴定过程正处于首次人体研究的关键路径上。我们严格检查了用于分离适合cGMP生产的细胞系的典型选择策略。在选择策略的不同评估阶段，对175个转基因进行了评估。鉴定出适合生产cGMP的高产细胞系。然而，我们的分析表明，最高产量细胞系的分离频率较低，并且每个评估阶段之间的排名位置不一致，这表明该策略有改进的潜力。通过对替代选择策略的计算机分析，试图增加10种最高产量细胞系的分离频率，但没有成功。我们确定了与典型选择策略具有相似预测能力的替代策略。一种替代策略需要在评估阶段进行较少的细胞系，但模型的随机性意味着细胞系数量可能在程序之间发生变化。总之，我们的研究阐明了改进这一和未来选择策略的潜力，基于使用更具信息性或减少方差的评估，为提高制造细胞系的产生效率铺平了道路。©2010美国化学工程师学会生物技术。掠夺。, 2010年
Transfectants with a wide range of cellular phenotypes are obtained during the process of cell line generation. For the successful manufacture of a therapeutic protein, a means is required to identify a cell line with desirable growth and productivity characteristics from this phenotypically wide‐ranging transfectant population. This identification process is on the critical path for first‐in‐human studies. We have stringently examined a typical selection strategy used to isolate cell lines suitable for cGMP manufacturing. One‐hundred and seventy‐five transfectants were evaluated as they progressed through the different assessment stages of the selection strategy. High producing cell lines, suitable for cGMP manufacturing, were identified. However, our analyses showed that the frequency of isolation of the highest producing cell lines was low and that ranking positions were not consistent between each assessment stage, suggesting that there is potential to improve upon the strategy. Attempts to increase the frequency of isolation of the 10 highest producing cell lines, by in silico analysis of alternative selection strategies, were unsuccessful. We identified alternative strategies with similar predictive capabilities to the typical selection strategy. One alternate strategy required fewer cell lines to be progressed at the assessment stages but the stochastic nature of the models means that cell line numbers are likely to change between programs. In summary, our studies illuminate the potential for improvement to this and future selection strategies, based around use of assessments that are more informative or that reduce variance, paving the way to improved efficiency of generation of manufacturing cell lines. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010