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包括TFREs在内的合成增强剂可以改善CHO细胞中的转基因表达

Hui-Ning Liu^1,2, Xiao-Yin Wang^1,3, Ying Zou^1,3

¹International Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, Xinxiang 453003, China.

|March 12, 2024

概括

合成增强剂通过优化细胞巨乳病毒 (CMV) 促进体,显著提高了中国仓鼠卵巢 (CHO) 细胞的重组蛋白质生产. 这一进展对于开发治疗性蛋白质的高产量CHO细胞表达系统至关重要.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
细胞生物学细胞生物学

背景情况:

人类细胞巨乳病毒主要的即时早期基因 (CMV) 促销者被广泛用于CHO细胞中的重组治疗蛋白 (RTP) 生产.
提高CHO细胞中的RTP产量对于高效的生物制药制造至关重要.

研究的目的:

评估合成增强剂是否能够增加CHO细胞中的重组蛋白水平.
研究这些增强剂对转基因表达和mRNA水平的影响.
探索增强剂介导基因表达背后的分子机制.

主要方法:

设计和测试了五种含有多重转录因子调节元件 (TFREs) 的合成增强剂.
报告者基因表达被评估在暂时和稳定转染的CHO细胞中.
人类血清白蛋白 (HSA) 的表达被用作模型治疗蛋白.
进行了转录学分析,以确定涉及的细胞通路.

主要成果:

五种合成增强剂中有四种在短暂和稳定转染中显著增加了记者基因表达.
HSA的短暂表达增加了1.5倍,而稳定表达增加了2.14倍.
增强的转基因表达与增加的mRNA水平相关.
转录组学数据表明转录激活和细胞循环调节途径的参与.

结论:

用合成增强剂优化CMV促进剂可以大幅增加CHO细胞中的转基因生产产量.
这些发现对开发更高效的CHO细胞基表达系统对生物治疗有重大影响.
已识别的增强剂为改善重组蛋白制造提供了一个有希望的策略.

关键词:

在CHO细胞表达系统中,在CMV中,CMV的宣传者是CMV的宣传者.增强器增强器增强器再组合蛋白质是一种重组蛋白质.合成促进剂合成促进剂

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