通过尺寸排阻色谱法从血浆中快速分离细胞外囊泡，随后进行基于质谱的蛋白质组学分析。
Rapid Isolation of Extracellular Vesicles from Blood Plasma with Size-Exclusion Chromatography Followed by Mass Spectrometry-Based Proteomic Profiling.
作者信息
Kreimer Simion, Ivanov Alexander R
机构信息
Department of Chemistry and Chemical Biology, Barnett Institute of Chemical and Biological Analysis, Northeastern University, 360 Huntington Avenue, 412TF, Boston, MA, 02115, USA.
出版信息
Methods Mol Biol. 2017;1660:295-302. doi: 10.1007/978-1-4939-7253-1_24.
Abstract
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.
摘要
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。
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本文引用的文献
1
Cancer: Diagnosis by extracellular vesicles.癌症：通过细胞外囊泡进行诊断。
Nature. 2015 Jul 9;523(7559):161-2. doi: 10.1038/nature14626. Epub 2015 Jun 24.
2
Mass-spectrometry-based molecular characterization of extracellular vesicles: lipidomics and proteomics.基于质谱的细胞外囊泡分子表征：脂质组学和蛋白质组学
J Proteome Res. 2015 Jun 5;14(6):2367-84. doi: 10.1021/pr501279t. Epub 2015 May 14.
3
Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles.最小实验要求定义细胞外囊泡及其功能：国际细胞外囊泡学会的立场声明。
J Extracell Vesicles. 2014 Dec 22;3:26913. doi: 10.3402/jev.v3.26913. eCollection 2014.
4
Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles.外泌体和其他细胞外囊泡的生物发生、分泌和细胞间相互作用。
Annu Rev Cell Dev Biol. 2014;30:255-89. doi: 10.1146/annurev-cellbio-101512-122326. Epub 2014 Aug 21.
5
Single-step isolation of extracellular vesicles by size-exclusion chromatography.通过大小排阻色谱法一步分离细胞外囊泡。
J Extracell Vesicles. 2014 Sep 8;3. doi: 10.3402/jev.v3.23430. eCollection 2014.
6
As we wait: coping with an imperfect nomenclature for extracellular vesicles.在等待的过程中：应对细胞外囊泡不完善的命名法。
J Extracell Vesicles. 2013 Feb 15;2. doi: 10.3402/jev.v2i0.20389. eCollection 2013.
7
Comparison of ultracentrifugation, density gradient separation, and immunoaffinity capture methods for isolating human colon cancer cell line LIM1863-derived exosomes.比较超速离心、密度梯度分离和免疫亲和捕获方法用于分离人结肠癌细胞系 LIM1863 来源的外泌体。
Methods. 2012 Feb;56(2):293-304. doi: 10.1016/j.ymeth.2012.01.002. Epub 2012 Jan 21.
8
A rapid method of total lipid extraction and purification.一种快速的总脂质提取与纯化方法。
Can J Biochem Physiol. 1959 Aug;37(8):911-7. doi: 10.1139/o59-099.
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