Efficient development of a stable cell pool for antibody production using a single plasmid
Efficient development of a stable cell pool for antibody production using a single plasmid
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使用单一质粒高效开发用于抗体生产的稳定细胞库
DOI：
10.1093/jb/mvy007
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发表时间：
2018-05
期刊：
影响因子：
--
通讯作者：
Xia Ningshao
中科院分区：
文献类型：
--
作者：
Yang Yi;You Min;Chen Fentian;Jia Tianrong;Chen Yuanzhi;Zhou Bin;Mi Qingyu;An Zhiqiang;Luo Wenxin;Xia Ningshao
治疗性抗体是增长最快的一类生物药物。候选药物的评价需要足够量的抗体。用稳定的细胞池生产抗体是生产用于药物候选物选择的蛋白质克数的有效策略。已经描述了许多用于开发用于抗体表达的稳定细胞库的方法。然而，大多数报道的方法是费力的，由于高生产者的频率低。在这项研究中，我们确定了最佳的载体和筛选参数，以开发一种有效构建稳定的抗体表达细胞池的策略。使用优化策略构建的细胞池始终产生更高的表达滴度，高达10倍的改善。此外，通过分子排阻色谱法评估，该方法导致具有高于95%的主要产物峰的细胞池的比例更高。通过对5个96孔板的筛选，可以获得高产菌株。该策略将大大减少临床电极导线选择期间的克隆筛选规模。该研究提供了一个平台，具有高效的质粒设计和筛选策略，用于在高表达双亚基蛋白质（如抗体）中显著节省成本和劳动力。
Therapeutic antibodies are the fastest growing group of biopharmaceuticals. Evaluation of drug candidates requires a sufficient amount of antibodies. Production of antibodies with stable cell pools is an efficient strategy to produce grams of proteins for drug candidate selection. Many methods have been described for developing stable cell pools for antibody expression. However, most of the reported methods are laborious due to the low frequency of high producers. In this study, we determined optimal vectors and screening parameters to develop a strategy for efficient construction of stable antibody expressing cell pools. The cell pool constructed using the optimized strategy consistently yielded a higher expression titer, up to 10-fold improvement. Further, this method resulted in a higher ratio of the cell pools with the main product peak above 95% as assessed by size-exclusion chromatography. High producers could be obtained by means of screening five 96-well plates. This strategy will greatly reduce clone-screening size during Clinical Lead Selection. This study provides a platform with efficient design of plasmids and screening strategies for significant cost and labour savings in high expression of two-subunit proteins such as antibodies.
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