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一个基于生物反应器的黄热病病毒样粒子生产工艺,采用基于短暂传染的综合工艺分析技术

Gregor Dekevic¹, Tobias Tertel², Lars Tasto¹

¹Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstrasse 14, 35390 Giessen, Germany.

|October 28, 2023

概括

开发一种类似黄热病疫苗的粒子 (YF-VLP) 生物反应器工艺,提供了一种新的被动免疫策略. 这种方法优化了等离子体DNA的产生和VLP的产量,以潜在地用于对抗被忽视的热带疾病.

科学领域:

生物技术是生物技术.
病毒学病毒学
生物工艺工程生物工艺工程

背景情况:

黄热病 (YF) 是一种严重的病毒性疾病,感染者没有快速治疗的选择.
使用YF病毒样颗粒 (YF-VLPs) 的被动免疫提供了一个潜在的治疗途径.
现有的幼年发烧预防仅依赖于疫苗接种,这凸显了对替代干预措施的需要.

研究的目的:

建立基于生物反应器的黄热病病毒样颗粒 (YF-VLPs) 生产工艺.
优化等离子体DNA (pDNA) 生产以提高VLP产量.
整合过程分析技术 (PAT) 进行实时过程监控和控制.

主要方法:

利用实验设计来优化pDNA生产,识别影响产量的关键媒体组件.
调整了HEK细胞密度,多重复形成时间和介质交换策略,以提高转染效率.
实施了一种混合生物反应器,集成介电光谱,用于实时监测细胞生长,多重复合吸收和收获时间.

主要成果:

实现了11mg/L的等离子体DNA产量,其中葡萄糖,NaCl,酵母提取物和酸盐缓冲剂显著影响了特定的pDNA产量.
通过优化生物反应器条件,证明了转染效率的提高,并将Pluronic F-68确定为中性和有害的抗凝聚剂.
通过Western blot,成像流细胞计和传输电子显微镜成功证实了YF-VLP的存在和完整性.

结论:

建立了一个强大的生物反应器过程,用于生产黄热病病毒样颗粒 (YF-VLPs).
开发的YF-VLP生产工艺可以被调整为一个平台,用于产生针对其他被忽视的热带疾病的VLP.
这项工作为开发针对病毒感染的被动免疫策略提供了基础.

关键词:

在HEK 293T细胞中.国际金融市场管理委员会 (IFCM)过程分析技术 (PAT) 是一种分析技术.设计实验的设计.这是一个很好的例子. lPEIEL.混合生物反应器 (STR)

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