Lippé Roger
Department of Pathology and Cell Biology, University of Montreal, V-541 Pavillon Roger Gaudry, 2900 boul. Édouard-Montpetit, Montreal, QC, Canada, H3C 3J7,
Methods Mol Biol. 2014;1144:181-90. doi: 10.1007/978-1-4939-0428-0_12.
The analysis of herpes simplex virus type 1 mature extracellular virions by proteomics requires highly enriched samples to limit false positives and favor the detection of true components. The protocol described below involves the removal of highly contaminating serum proteins and purification of the virions by a series of differential and density centrifugation steps. In addition, L-particles, which are viral particles devoid of genome and capsid but present in the extracellular milieu, are depleted on Ficoll 400 gradients. As previously reported, the resulting viral particles are free of most contaminants and suitable for mass spectrometry.
通过蛋白质组学分析1型单纯疱疹病毒成熟的细胞外病毒粒子需要高度富集的样本，以限制假阳性并有利于检测真正的成分。以下所述方案包括去除高度污染的血清蛋白，并通过一系列差速离心和密度离心步骤纯化病毒粒子。此外，L颗粒（即缺乏基因组和衣壳但存在于细胞外环境中的病毒颗粒）在Ficoll 400梯度上被去除。如先前报道的那样，所得病毒颗粒不含大多数污染物，适用于质谱分析。
