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首页> 外文期刊>Biotechnology Progress >Sustaining an efficient and effective CHO cell line development platform by incorporation of 24-deep well plate screening and multivariate analysis
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Sustaining an efficient and effective CHO cell line development platform by incorporation of 24-deep well plate screening and multivariate analysis

机译：通过掺入24深井板筛选和多变量分析来维持高效且有效的CHO细胞系开发平台

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摘要

Efficient and effective cell line screening is paramount toward a successful biomanufacturing program. Here we describe the implementation of 24-deep well plate (24-DWP) screening of CHO lines as part of the cell line development platform at AbbVie. Incorporation of this approach accelerated the identification of the best candidate lines for process development. In an effort to quantify and predict process performance comparability, we compared cell culture performance in and in shake flasks, for a panel of Chinese Hamster Ovary cell lines expressing a monoclonal antibody. The results in 24-DWP screening showed reduced growth profiles, but comparable viability profiles. Slow growers in 24-DWP achieved the highest productivity improvement upon scaling-up to shake flasks. Product quality of the protein purified from shake flasks and 24-DWP were also compared. The 24-DWP culture conditions were found to influence the levels of acidic species, reduce the G0 N-glycan species, and increase the high-mannose N-glycan species. Nevertheless, the identification of undesirable profiles is executed consistently with the scaled-up culture. We further employed multivariate data analysis to capture differences depending on the two scales and we could demonstrate that cell line profiles were adequately clustered, regardless of the vessel used for the development. In conclusion, the 24-DWP platform was reasonably predictive of the parameters crucial for upstream process development activities, and has been adapted as part of the AbbVie cell line development platform. (c) 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:175-186, 2018
机译：高效且有效的细胞系筛选对于成功的生物制造计划至关重要。在这里，我们描述了24深井板（24-DWP）筛选CHO系列作为ABBVIE的细胞系开发平台的一部分。本方法的纳入加速了用于过程开发的最佳候选线路的识别。为了量化和预测过程性能可比性，我们对表达单克隆抗体的中国仓鼠卵巢细胞系小组进行了比较细胞培养性能和摇动烧瓶中的细胞培养性能。结果在24-DWP筛选中显示出增长型材的降低，但可比较的可行性轮廓。 24-DWP中的慢种植者在扩大速度震动烧瓶时取得了最高的生产率改善。还比较了从摇瓶和24-DWP纯化的蛋白质的产品质量。发现24-DWP培养条件影响酸性物质的水平，降低G0 N-聚糖物种，并增加高甘露糖N-聚糖物种。然而，用缩放培养始终执行不希望的曲线的识别。我们进一步采用多变量数据分析来捕获根据两种尺度的差异，我们可以证明细胞线轮廓是充分聚集的，无论用于开发的血管如何。总之，24-DWP平台合理预测上游工艺开发活动至关重要的参数，并被调整为ABBVIE细胞系开发平台的一部分。 （c）2017美国化学工程师生物科技学院。 Prog。，34：175-186,2018

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