在选择过程中抑制难以表达的重组蛋白的表达,提高了CHO稳定池的生产力
Jean-Sébastien Maltais1, Simon Lord-Dufour1, Audrey Morasse1
1Mammalian Cell Expression, Human Health Therapeutics Research Centre, National Research Council Canada, Montréal, Québec, Canada.
Biotechnology and bioengineering
|May 26, 2023
概括
在选择过程中使用可诱导的中国子卵巢 (CHO) 系统来减少重组蛋白 (r-蛋白) 表达,可以提高细胞池的生产力. 这种方法最大限度地减少了细胞应激,从而提高了治疗性蛋白质的产量.
科学领域:
- 生物技术是生物技术.
- 细胞生物学 细胞生物学
- 蛋白质工程是指蛋白质的工程.
背景情况:
- 中国仓鼠卵巢 (CHO) 细胞是复合蛋白 (r-蛋白) 制造的标准.
- 构成性表达系统面临的挑战是复杂的治疗方法,如细胞因子和双特异性抗体.
研究的目的:
- 为了评估一个可化诱导的CHO平台,以改善r蛋白的生产.
- 为了比较带有和没有感应选择的池的生产率.
主要方法:
- 开发了一种可化诱导的CHO表达系统.
- 在选择过程中减少r-蛋白表达 (OFF-pool) 和构成性表达 (ON-pool) 的稳定细胞池生成.
- 进行了料批量生产,以评估各种r蛋白的体积生产率.
主要成果:
- 在10个测试的r蛋白中,OFF池对8个r蛋白的生产率明显高于ON池.
- OFF池显示出更高比例的高表达细胞,当表达关闭时,它们的增殖速度更快.
- 在OFF池中观察到细胞应激减少 (ER应激,代谢负担),与更高的生产率相关.
结论:
- 在稳定的CHO池选择过程中通过可诱导系统最小化r-蛋白表达,可以提高细胞池质量.
- 这一策略可以提高各种治疗性蛋白质的体积生产率.
- 诱导性系统减轻了代谢负担和ER压力,使得产生更强壮,高生产率的细胞系成为可能.
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