Proteomic studies of circulating vesicles are hampered by difficulties in purifying vesicles from plasma and serum. Isolations are contaminated with high-abundance blood proteins that may mask genuine vesicular-associated proteins and/or simply provide misleading data. In this brief report, we explored the potential utility of a commercially available size exclusion chromatography column for rapid vesicle purification. We evaluated the performance of the column, with cancer cell line conditioned medium or healthy donor plasma, in terms of removing non-vesicular protein and enriching for vesicles exhibiting exosome characteristics. Serial fractions revealed a peak for typical exosomal proteins (CD9, CD81 etc.) that preceded the peak for highly abundant proteins, including albumin, for either sample type, and harvesting only this peak would represent elimination of >95% of protein from the sample. The columns showed good reproducibility, and streamlining the workflow would allow the exosome-relevant material to be collected in less than 10 minutes. Surprisingly, however, subsequent post-column vesicle concentration steps whilst resulting in some protein loss also lead to low vesicle recoveries, with a net effect of reducing sample purity (assessed by the particle-to-protein ratio). The columns provide a convenient, reproducible and highly effective means of eliminating >95% of non-vesicular protein from biological fluid samples such as plasma.

循环囊泡的蛋白质组学研究因难以从血浆和血清中纯化囊泡而受到阻碍。分离物被高丰度的血液蛋白污染，可能掩盖真正的囊泡相关蛋白和/或只是提供误导性数据。在这份简短的报告中，我们探讨了市售尺寸排阻色谱柱在快速囊泡纯化中的潜在用途。我们使用癌细胞系条件培养基或健康供体血浆评估了该色谱柱在去除非囊泡蛋白和富集表现出外泌体特征的囊泡方面的性能。系列级分显示典型外泌体蛋白（CD9、CD81 等）的峰值在高丰度蛋白质（包括白蛋白）的峰值之前，对于任一样品类型，并且仅收获该峰将代表从样品中消除> 95％的蛋白质。这些色谱柱显示出良好的重现性，简化工作流程将允许在不到 10 分钟的时间内收集外泌体相关材料。然而，令人惊讶的是，随后的柱后囊泡浓缩步骤在导致一些蛋白质损失的同时也导致了低囊泡回收率，净效应降低了样品纯度（通过颗粒与蛋白质的比率评估）。该色谱柱提供了一种方便、可重复且高效的方法，可从血浆等生物流体样品中去除 >95% 的非囊泡蛋白。简化工作流程将允许在不到 10 分钟的时间内收集外泌体相关材料。然而，令人惊讶的是，随后的柱后囊泡浓缩步骤在导致一些蛋白质损失的同时也导致了低囊泡回收率，净效应降低了样品纯度（通过颗粒与蛋白质的比率评估）。该色谱柱提供了一种方便、可重复且高效的方法，可从血浆等生物流体样品中去除 >95% 的非囊泡蛋白。简化工作流程将允许在不到 10 分钟的时间内收集外泌体相关材料。然而，令人惊讶的是，随后的柱后囊泡浓缩步骤虽然会导致一些蛋白质损失，但也会导致囊泡回收率低，净效应会降低样品纯度（通过颗粒与蛋白质的比率评估）。该色谱柱提供了一种方便、可重复且高效的方法，可从血浆等生物流体样品中去除 >95% 的非囊泡蛋白。