Automation of cell line development
Automation of cell line development
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DOI：
10.1007/s10616-009-9187-y
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发表时间：
2009-01-01
期刊：
影响因子：
2.2
通讯作者：
Skoging-Nyberg, Ulrica
中科院分区：
文献类型：
--
作者：
Lindgren, Kristina;Salmen, Andrea;Skoging-Nyberg, Ulrica
已经建立了用于开发用于生物制药生产的高产细胞系的自动化平台，以增加产量并降低开发成本。该概念基于Cello机器人系统（The Automation Partnership），涵盖菌落筛选和静态培养物扩增。在本研究中，用于在中国仓鼠卵巢细胞中生产治疗性单克隆抗体的谷氨酰胺合成酶表达系统（Lonza Biologics）用于评价自动化方法。结果表明，在转染后的菌落检测和筛选步骤中IgG滴度的分布方面，自动化程序能够产生与传统产生的细胞系质量相同的细胞系。在搅拌罐生物反应器中的通用补料分批评价中，获得了最佳表达细胞系的4.7和5.0 g/L的IgG滴度。我们估计，与手动过程相比，使用自动化过程完成的细胞系开发项目的数量可以增加多达三倍，而不会增加手动工作量。发现早期筛选中获得的IgG滴度与摇瓶中分批补料培养中获得的滴度之间的相关性较差。这进一步暗示了利用能够筛选和扩增大量转染子的高通量系统的益处。应用两种浓度，56和75 μ M的选择剂甲硫氨酸磺酰亚胺（MSX），以评价对转染后获得的集落数的影响。当应用含有75 μ M MSX的选择培养基时，与用56 μ M MSX选择的细胞系相比，获得较少的低产量转染子，但发现相同数量的高产细胞系。
An automated platform for development of high producing cell lines for biopharmaceutical production has been established in order to increase throughput and reduce development costs. The concept is based on the Cello robotic system (The Automation Partnership) and covers screening for colonies and expansion of static cultures. In this study, the glutamine synthetase expression system (Lonza Biologics) for production of therapeutic monoclonal antibodies in Chinese hamster ovary cells was used for evaluation of the automation approach. It is shown that the automated procedure is capable of producing cell lines of equal quality to the traditionally generated cell lines in terms of colony detection following transfection and distribution of IgG titer in the screening steps. In a generic fed-batch evaluation in stirred tank bioreactors, IgG titers of 4.7 and 5.0 g/L were obtained for best expressing cell lines. We have estimated that the number of completed cell line development projects can be increased up to three times using the automated process without increasing manual workload, compared to the manual process. Correlation between IgG titers obtained in early screens and titers achieved in fed-batch cultures in shake flasks was found to be poor. This further implies the benefits of utilizing a high throughput system capable of screening and expanding a high number of transfectants. Two concentrations, 56 and 75 mu M, of selection agent, methionine sulphoximine (MSX), were applied to evaluate the impact on the number of colonies obtained post transfection. When applying selection medium containing 75 mu M MSX, fewer low producing transfectants were obtained, compared to cell lines selected with 56 mu M MSX, but an equal number of high producing cell lines were found. By using the higher MSX concentration, the number of cell line development projects run in parallel could be increased and thereby increasing the overall capacity of the automated platform process.