超越氨酸养:一种基于重组CHO细胞中氨酸代谢工程的新型食批量培养策略
Lei Cao1, Liang Zhao2, Qian Ye3
1State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China; Shanghai BioEngine Sci-Tech Co., LTD, Shanghai 201203, China.
Journal of biotechnology
|October 12, 2025
概括
这项研究优化了CHO细胞生物制造中的氨酸供应,通过与现有的酶共同过度表达类二二二缩酶 (QDPR). 这种新的策略使单克隆抗体的产生增加了30%以上,而没有复杂的铁素料.
科学领域:
- 生物技术是生物技术.
- 生物工艺工程 生物工艺工程
- 代谢工程是代谢工程.
背景情况:
- 中国仓鼠卵巢 (CHO) 细胞对于单克隆抗体 (mAb) 生产至关重要.
- 有限的氨酸溶解度和复杂的养策略阻碍了高密度养批次培养.
- 以前的努力过度表达了丁-4α-碳胺脱水酶1 (PCBD1) 和氨酸氧酶 (PAH),但并没有完全解决供应问题.
研究的目的:
- 在重组CHO (rCHO) 细胞中识别和解决内源性氨酸合成的瓶.
- 开发一个改进的料批量策略,以提高高密度培养中的mAb生产.
- 通过消除具有挑战性的性铁素料来简化营养需求.
主要方法:
- 验证了rCHO细胞对外源性氨酸的依赖.
- 分析了PCBD1,氨酸氧化酶 (PAH) 和酸二二二缩酶 (QDPR) 的表达水平.
- 与PCBD1和PAH一起共同过度表达QDPR,以重塑氨酸生物合成途径.
- 实施了一种新的养批量策略,使用基底介质含有3.0mM氨酸和无氨酸的单次养介质.
主要成果:
- 证实低PCBD1/PAH水平限制了内源性氨酸合成.
- 鉴于QDPR在氨酸有限的条件下被确定为一个关键瓶.
- 新的食批量策略实现了4.24g/L的最终mAb标位.
- 这比仅过度表达PCBD1/PAH的细胞增加了32.50%,比传统方法增加了10.70%.
结论:
- 包括QDPR在内的整体代谢途径优化对于高效的生物制造至关重要.
- 开发的策略简化了营养供应,通过消除性氨酸料.
- 这种方法为基于CHO细胞的高密度,氨酸有限的mAb生产提供了显著的价值.
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