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通过在复合性CHO细胞基础的短暂基因表达系统中的过程优化,提高etanercept生产中的蛋白质纯度

Hyun-Seung Kim^1,2, Hoon-Min Lee¹, Tae-Ho Kim^1,3

¹Biotherapeutics Translational Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, Korea.

Bioprocess and biosystems engineering

|March 9, 2026

概括

优化短暂基因表达 (TGE) 条件,包括拉巴素治疗和温度变化,显著提高治疗性葡萄糖蛋白的纯度. 这些增强可以达到与稳定基因表达 (SGE) 系统可比的纯度水平.

科学领域:

生物技术是生物技术.
蛋白质生产蛋白质生产
哺乳动物细胞培养

背景情况:

高蛋白质纯度对于治疗性葡萄糖蛋白的有效性和安全性至关重要.
暂时基因表达 (TGE) 系统是用于治疗性蛋白质生产的稳定基因表达 (SGE) 系统的前身.
评估和提高TGE中的蛋白质纯度对于下游应用至关重要.

研究的目的:

评估和提高通过TGE生产的乙素 (ETN) 的纯度.
为了比较TGE和SGE系统之间的蛋白质纯度.
调查过程修改,以提高TGE中的ETN纯度.

主要方法:

在过渡性基因表达 (TGE) 系统中评估了乙素 (ETN) 产量.
工艺修改包括化学处理 (拉帕米辛),温度降低 (30°C),以及对套件组件的调整 (Enhancer遗漏).
通过分析单体比例和分子量杂质来评估蛋白质纯度.

主要成果:

与SGE相比,TGE生产的ETN表现出较高的高分子和低分子重量的杂质.
拉帕米辛治疗,30°C的温度下降,以及Enhancer的缺失增加了ETN单体的比例.
结合优化条件 (拉帕米辛,30°C,延长培养) 显著提高了ETN的生产和纯度,达到与SGE可比的水平.

结论:

优化TGE培养参数可以显著提高治疗性糖蛋白纯度.
在TGE中实现的纯度水平与SGE系统中的纯度水平相似.
优化的条件显示了哺乳动物TGE系统的广泛适用性,包括HEK293细胞.

关键词:

在Etanercept之外的其他地方人类胚胎脏293 (HEK293) 细胞蛋白质的纯度蛋白质的纯度再组合的中国仓鼠卵巢 (CHO) 细胞.暂时的基因表达.

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