Asymmetric flow field flow fractionation methods for virus purification
Asymmetric flow field flow fractionation methods for virus purification
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DOI：
10.1016/j.chroma.2016.09.055
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发表时间：
2016-10-21
影响因子：
4.1
通讯作者：
Oksanen, Hanna M.
中科院分区：
文献类型：
--
作者：
Eskelin, Katri;Lampi, Mirka;Oksanen, Hanna M.
病毒的详细生化和生物物理特征需要高质量和高纯度的病毒制剂。优化病毒的生产和纯化是一个必不可少的过程，但既费力又耗时。非对称流分离(AF4)是一种快速、温和的病毒分离方法，可以保持病毒的感染性，是一种很有吸引力的病毒分离方法。在这里，我们优化了AF4条件，以用于从各种类型的起始材料中纯化模型病毒--噬菌体PRD1。我们的结果表明，从病毒回收率和特异性感染性来看，AF4非常适合于PRD1的纯化。较短的分析时间和较高的进样量使我们能够使用AF4进行PRD1的制备规模纯化。此外，我们证明AF4能够快速实时分析感染细胞中的子代病毒生产。(C)2016爱思唯尔B.V.保留所有权利。
Detailed biochemical and biophysical characterization of viruses requires viral preparations of high quantity and purity. The optimization of virus production and purification is an essential, but laborious and time-consuming process. Asymmetric flow field flow fractionation (AF4) is an attractive alternative method for virus purification because it is a rapid and gentle separation method that should preserve viral infectivity. Here we optimized the AF4 conditions to be used for purification of a model virus, bacteriophage PRD1, from various types of starting materials. Our results show that AF4 is well suited for PRD1 purification as monitored by virus recovery and specific infectivity. Short analysis time and high sample loads enabled us to use AF4 for preparative scale purification of PRD1. Furthermore, we show that AF4 enables the rapid real-time analysis of progeny virus production in infected cells. (C) 2016 Elsevier B.V. All rights reserved.