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生物质密集型CHO细胞的定向进化通过适应亚生理温度

Katie L Syddall¹, Alejandro Fernandez-Martell¹, Joseph F Cartwright¹

¹Department of Chemical and Biological Engineering, University of Sheffield, Mappin St., Sheffield, S1 3JD, UK.

Metabolic engineering

|November 25, 2023

概括

在32°C下对150代中国仓鼠卵巢 (CHO) 细胞的定向进化增强了细胞生长,并显著提高了重组蛋白质的产生. 适应寒冷的CHO细胞表现出增加的生物质,线粒体功能和抗氧化能力,证明了细胞工程的基因组规模可塑性.

科学领域:

生物技术是生物技术.
细胞生物学细胞生物学
基因组学就是基因组学.

背景情况:

中国仓鼠卵巢 (CHO) 细胞被广泛用于重组蛋白质生产.
提高宿主细胞的生物合成能力对于生物制药制造至关重要.
细胞适应环境条件可以提高生产力.

研究的目的:

开发一种方法来提高宿主CHO细胞的生物合成能力.
为了研究长时间的亚生理温度培养对CHO细胞特征的影响.
探索指导进化作为CHO细胞工程的战略.

主要方法:

在Lonza专有CHOK1SV®细胞的系列亚种植中,在32°C下进行了150多代的生产.
监测细胞增殖率和表型特征.
分析细胞体积,生物质含量 (RNA,蛋白质),线粒体功能,抗氧化能力和中心代谢.
评估重组蛋白 (IgG4单克隆抗体,Fc融合蛋白) 的生产力和N-甘氨酸加工.
对型和mRNA丰度的基因组规模分析.

主要成果:

低温培养导致了与37°C培养相比的繁殖率的恢复.
适应寒冷的细胞体积增加,生物量增加,线粒体功能增加,抗氧化能力增加.
中央新陈代谢发生了改变,而N-甘氨酸加工没有受到影响.
重组蛋白的生产力 (IgG4 mAb,Fc融合) 显著增加.
观察到型和mRNA丰度的显著基因组规模变化.

结论:

32°C的定向进化是一种有效的策略,可以提高CHO细胞的生物合成能力.
对亚生理温度的协调细胞适应导致蛋白质生产的改善.
CHO细胞具有显著的基因组和功能性可塑性,可用于细胞工程.

关键词:

在CHO细胞中.定向进化是指导进化的.代谢工程是代谢工程.单克隆抗体是一种单克隆抗体.重组疗法药物重组疗法药物

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