从Pichia pastoris使用响应表面方法和人工神经网络来建模和优化复合托西利祖马布生产
Prabir Kumar Das1, Ansuman Sahoo1, Venkata Dasu Veeranki1
1Biochemical Engineering Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, India.
Biotechnology and bioengineering
|May 15, 2025
概括
在Pichia pastoris中优化定义介质显著提高了重组单克隆抗体 (mAb) 生产. 曼尼托和甘氨酸增强了全长的托西利祖马布 (TCZ),而和硫酸盐提高了Fab碎片的产量.
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
- 分子生物学分子生物学
背景情况:
- 牧羊是用于重组蛋白质生产的广泛使用的宿主,包括单克隆抗体 (mAbs).
- 优化定义媒体对于最大限度地提高mAb产量和确保产品的一致性至关重要.
- 托西利祖马布 (TCZ) 是一种具有重要的临床应用的治疗药物.
研究的目的:
- 在Pichia pastoris中优化一个定义的介质,以增强复合全长托西利祖马布 (TCZ) 和其Fab片段的生产.
- 确定影响TCZ和Fab生产的关键介质组件及其最佳水平.
- 评估统计和计算方法在预测和改善mAb产量的有效性.
主要方法:
- 对四种定义的介质进行选,然后确定最佳的碳 (曼尼托,) 和 (甘氨酸,硫酸盐) 来源.
- 应用Plackett-Burman设计来识别影响TCZ和Fab生产的重要介质组件.
- 使用响应表面方法 (盒式设计) 和人工神经网络 (ANN) 与遗传算法 (GA) 相结合的选组件的优化.
主要成果:
- 发现FM22定义的介质适合复合菌株生长和抗体产量.
- 确定了最佳的碳和来源:全长TCZ的曼尼托尔/甘氨酸和Fab片段的酸盐/酸盐.
- 优化的介质组件导致震动瓶产量为0.35 mg/L (全长TCZ) 和0.42 g/L (Fab),批量反应堆产量为0.44 mg/L (全长TCZ) 和0.45 g/L (Fab).
- 实现了全长TCZ的3.8倍和Fab的2.9倍的整体产量增加.
结论:
- 定义的介质优化显著提高了Pichia pastoris.的重组mAb产量.
- 普莱克特-伯曼,盒子-贝恩肯设计,ANN和GA的结合为生物过程优化提供了一个强大的方法.
- 优化过程表明,托西利祖马布及其Fab片段的产量得到了大幅改善,为更高效的生物制药制造铺平了道路.
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