在HEK293细胞中优化了rhFVII的生产和凝固活性,用于治疗出血障碍
Sen Zou1, Xiaoxiao Li2, Jiajun Liu2
1NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100050, China.
Protein expression and purification
|December 5, 2025
概括
研究人员优化了HEK293细胞中复合人体凝血因子VII (rhFVII) 的产生. 这一进步为改善生物制药制造和更好地治疗血友病等出血障碍提供了潜力.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 血友病是一种遗传出血疾病,需要有效的治疗方法.
- 重组凝血因子对于血友病治疗至关重要.
- 优化表达系统是高效生物制药生产的关键.
研究的目的:
- 为了优化HEK293细胞中复合人体凝血因子VII (rhFVII) 的表达.
- 为了提高rhFVII在治疗出血障碍方面的疗效.
- 为大规模生产建立稳定的细胞线. rhFVII生产.
主要方法:
- 将人类FVII基因克隆到pcDNA3.1载体中.
- 优化暂时转染条件 (DNA:PEI比率,细胞密度).
- 选择和确认稳定的FVII表达细胞系 (G418,RT-PCR) 并适应悬浮培养.
- 分析rhFVII表达 (SDS-PAGE,西部斑块,ELISA) 和凝血活动 (前列红素时间测试).
主要成果:
- 过渡性转染实现了rhFVII度为10.22μg/L,凝血活性为232.46%.
- 稳定的HEK293细胞系在悬浮中产生11.22 mg/L的rhFVII.
- 最大凝血活性达到了563.17%,证明了高稳定性和安全性.
结论:
- 在HEK293细胞中成功优化了rhFVII表达,以增强生物制药生产.
- 开发出稳定,高产的细胞系,以产生功能性的rhFVII.
- 这项工作为改善血友病治疗和生物制造工艺铺平了道路.
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