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采用磁硫酸化纤维素颗粒与已建立的离心分析法比较流感病毒粒子的纯化。

Comparison of Influenza Virus Particle Purification Using Magnetic Sulfated Cellulose Particles with an Established Centrifugation Method for Analytics.

机构信息

Max Planck Institute for Dynamics of Complex Technical Systems , Sandtorstrasse 1, 39106 Magdeburg, Germany.

Otto von Guericke University Magdeburg , Universitätsplatz 2, 39106 Magdeburg, Germany.

出版信息

Anal Chem. 2015 Nov 3;87(21):10708-11. doi: 10.1021/acs.analchem.5b02681. Epub 2015 Oct 14.

Abstract

A method for the purification of influenza virus particles using novel magnetic sulfated cellulose particles is presented and compared to an established centrifugation method for analytics. Therefore, purified influenza A virus particles from adherent and suspension MDCK host cell lines were characterized on the protein level with mass spectrometry to compare the viral and residual host cell proteins. Both methods allowed one to identify all 10 influenza A virus proteins, including low-abundance proteins like the matrix protein 2 and nonstructural protein 1, with a similar impurity level of host cell proteins. Compared to the centrifugation method, use of the novel magnetic sulfated cellulose particles reduced the influenza A virus particle purification time from 3.5 h to 30 min before mass spectrometry analysis.

摘要

介绍了一种使用新型磁性硫酸化纤维素颗粒纯化流感病毒颗粒的方法,并将其与用于分析的既定离心方法进行了比较。因此,用质谱法对贴壁和悬浮 MDCK 宿主细胞系中纯化的甲型流感病毒颗粒进行了蛋白质水平的表征,以比较病毒和残留宿主细胞蛋白。两种方法都可以鉴定出所有 10 种甲型流感病毒蛋白,包括丰度较低的蛋白,如基质蛋白 2 和非结构蛋白 1,同时宿主细胞蛋白的杂质水平也相似。与离心方法相比,在进行质谱分析之前,使用新型磁性硫酸化纤维素颗粒将甲型流感病毒颗粒的纯化时间从 3.5 小时缩短至 30 分钟。

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