通过响应表面方法和微生物反应器查优化再生微型胰岛素的生产
Esra Ayan1,2,3, Ali Özhan Aytekin4, Ahmet Kati2,3
1Department of Molecular Biology and Genetics, Faculty of Science, Koç University, Istanbul, Türkiye.
PloS one
|September 8, 2025
概括
通过混合微尺度和统计建模方法进行工程化微型胰岛素 (nMPI) 生产. 优化的生物处理实现了高产量,证明了E. coli重组蛋白表达的可扩展平台.
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:
- 再组合胰岛素的生产需要可扩展的,高产量的,具有成本效益的生物工艺.
- 新型迷你亲胰岛素 (nMPI) 设计用于增强表达和简化切割.
- 优化nMPI生产对于满足不断增长的需求至关重要.
研究的目的:
- 开发和优化一种高产量,可扩展的生物工艺,用于新型微型胰岛素 (nMPI) 生产.
- 应用一种混合方法,将微量种植和统计建模相结合,以优化流程.
- 验证开发的优化系统的预测准确性和可扩展性.
主要方法:
- 经过改造的C-和残留物替代物的新型微型胰岛素 (nMPI).
- 使用的BioLector微生物反应器用于微尺度高吞吐量培养.
- 采用响应表面方法 (RSM),包括Plackett-Burman设计 (PBD) 和中央复合设计 (CDD),用于媒介优化.
- 将优化过程扩展到3L生物反应器.
主要成果:
- 鉴定出甘油是对nMPI产量最有影响力的介质成分.
- 优化的介质配方 (情景III) 在微观种植中实现了13.00 g/L的生产率.
- 在扩大规模时保持高性能,在3L生物反应器中达到11.5g/L.
- 已证明平衡的碳和来源增强了细胞活力和蛋白质表达.
结论:
- 混合优化系统准确地预测并有效地扩展.
- 为大肠杆菌中的nMPI建立了一个强大的和可扩展的生产平台.
- 提出的工作流作为高效的重组蛋白表达系统的模型.
- 开发的生物工艺适合转化为工业环境.
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