文献类型：期刊文献

中文题名：基于PAT的PCV2 VLPs生产过程优化与控制研究

英文题名：Process Control and Optimization of PCV2 Production of VLPs Based on PAT

作者：化磊召[1];易小萍[1];储炬[1];庄英萍[1];张嗣良[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室

年份：2018

卷号：38

期号：8

起止页码：50

中文期刊名：中国生物工程杂志

外文期刊名：China Biotechnology

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2017_2018】；

语种：中文

中文关键词：VLPs;昆虫细胞-杆状病毒载体表达系统;PAT

外文关键词：VLPs;BVES;PAT

摘要：昆虫细胞-杆状病毒载体表达系统（baculovirus expression vector system,BEVS）是病毒样颗粒亚单位疫苗（virus like particles,VLPs）的理想生产平台。动物细胞培养过程分析技术（process analytical technology,PAT）研究的重点在于更多地获取与细胞生理状态相关的过程参数,以及实现过程敏感参数的在线表征和过程控制关键时间节点的在线判定,从而指导过程优化和控制。通过昆虫Sf9细胞的分批和补料悬浮培养,发现细胞代谢活性与在线特征频率（fc）之间存在相关性。以fc作为细胞代谢活性的在线指征,在细胞代谢活性下降之前补料,使得Sf9细胞在代谢活性明显增强的基础上,最高活细胞密度提高1.75倍。通过分批培养与补料分批培养过程细胞生理特性参数的相关性分析,发现比电容增长速率（με）与培养体系S-期细胞比例之间存在相关性,με作为细胞增殖活性状态的在线指征参数,可以作为最佳接毒时间的判定依据。此外,研究还发现在线检测参数电容（ε）与最高疫苗产量之间存在相关性,可以作为疫苗最佳收获时间的在线表征参数。以在线fc值作为补料时间指征,以在线με值作为病毒感染时间指征,以在线ε值作为病毒收获时间指征,实现了猪圆环病毒2型（PCV2）VLPs的高效生产。相比于批培养,基于PAT的生产工艺疫苗单位体积产量提高76%,生产周期缩短了24h,为PCV2病毒样颗粒亚单位疫苗大规模生产提供了一种新的高效生产模式。
Baculovirus expression vector system（ BEVS） is the ideal platform for Virus-Like Particles（ VLPs） production. The key points of the Process Analytical Technology（ PAT） in animal cell culture include obtaining cell physiological characteristic parameters,monitoring process sensitive parameters,and controlling critical time points. These parameters in turn can promote process optimization and process control. A close correlation between cell metabolic status and on-line parameter critical frequency（ fc） was found in Sf 9 cell batch culture. Using the fcas on-line indictor of cell metabolic status and feeding the culture,a 1. 7 time increase of Peak Viable Cell Density（ PVCD） was obtained. Besides,the metabolic status was also improved. A close correlation was found between the proportion of S-phase cells and specific capacitance growth rate（ με）. It turned out that μεcould indicate the optimized infection time point. Additionally,a high correlation was found betweenε and the maximum VLPs productivity. The parameter of ε can act as an on-line indicator. Overall,an effective product mode was established by using fcas on-line feeding indicator,μεas on-line infecting indicator,ε as online harvesting indicator. Compared with traditional process,the volume productivity was increased by 76%,and the culture duration was reduced by 24 h. A new VLPs production mode has been established successfully.