A generalized purification step for viral particles using mannitol flocculation
A generalized purification step for viral particles using mannitol flocculation
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DOI：
10.1002/btpr.2651
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发表时间：
2018-07
影响因子：
2.9
通讯作者：
C. Heldt;Ashish Saksule;Pratik U. Joshi;M. Ghafarian
中科院分区：
文献类型：
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作者：
C. Heldt;Ashish Saksule;Pratik U. Joshi;M. Ghafarian
疫苗生产通常由发达国家使用传统的单元操作如过滤和色谱法进行。目前，疫苗的生产正在向欠发达国家转移，这就需要降低成本、提高回收率并适合连续生产的单元操作。这项工作表明，甘露醇可用作有包膜和无包膜病毒的絮凝剂，并可在微滤后从蛋白质污染物中纯化病毒。病毒的回收率范围为58 - 96%，取决于病毒、过滤器孔径和病毒的起始浓度。使用0.1 µm过滤器，小型无包膜病毒的蛋白质去除率达到80%，因为蛋白质不会与病毒絮凝并流过过滤器。假设甘露醇通过控制病毒周围的水结构使病毒表面脱水。由于病毒没有蛋白质的压缩能力，因此在渗透剂存在下病毒会聚集，而蛋白质则不会。渗透剂絮凝是一种使用高通量微滤器的可扩展工艺。它已被应用于包膜和无包膜病毒，使这一过程对各种疫苗和基因治疗产品友好。© 2018美国化学工程师学会Biotechnol.程序：34：1027-1035，2018
Vaccine manufacturing has conventionally been performed by the developed world using traditional unit operations like filtration and chromatography. There is currently a shift in the manufacturing of vaccines to the less developed world, requiring unit operations that reduce costs, increase recovery, and are amenable to continuous manufacturing. This work demonstrates that mannitol can be used as a flocculant for an enveloped and nonenveloped virus and can purify the virus from protein contaminants after microfiltration. The recovery of the virus ranges from 58 to 96% depending on virus, the filter pore size, and the starting concentration of the virus. Protein removal of 80% was achieved for the small nonenveloped virus using a 0.1 µm filter because proteins were not flocculated with the virus and flowed through the filter. It is hypothesized that mannitol dehydrates the viral surface by controlling the water structure surrounding the virus. Without the ability to become compact, as occurs with proteins, the virus aggregates in the presence of osmolytes and proteins do not. Osmolyte flocculation is a scalable process using high flux microfilters. It has been applied to both an enveloped and nonenveloped virus, making this process friendly to a variety of vaccine and gene therapy products. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1027–1035, 2018