Kreimer Simion, Ivanov Alexander R
Department of Chemistry and Chemical Biology, Barnett Institute of Chemical and Biological Analysis, Northeastern University, 360 Huntington Avenue, 412TF, Boston, MA, 02115, USA.
Methods Mol Biol. 2017;1660:295-302. doi: 10.1007/978-1-4939-7253-1_24.
The presented procedure allows rapid isolation of extracellular vesicles (EVs) from plasma using size-exclusion chromatography (SEC). Additionally, an approach for reducing the lipid and salt content of the EV isolate in preparation for mass spectrometry (MS)-based proteomic analysis is presented. An example setup for proteomic profiling of the processed samples by nanoflow liquid chromatography coupled to tandem mass spectrometry (nLC-MS/MS) is also presented. Approximately 1000 protein groups in blood plasma-derived EVs can be identified and quantitated following this procedure and using the described instrumentation.
所提出的方法允许使用尺寸排阻色谱法（SEC）从血浆中快速分离细胞外囊泡（EVs）。此外，还提出了一种在为基于质谱（MS）的蛋白质组学分析做准备时降低EV分离物中脂质和盐含量的方法。还给出了通过纳流液相色谱-串联质谱（nLC-MS/MS）对处理后的样品进行蛋白质组学分析的示例设置。按照此方法并使用所描述的仪器，可鉴定和定量血浆来源的EVs中的约1000个蛋白质组。
