Detailed analysis of the plasma extracellular vesicle proteome after separation from lipoproteins.
Detailed analysis of the plasma extracellular vesicle proteome after separation from lipoproteins.
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DOI：
10.1007/s00018-018-2773-4
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发表时间：
2018-08
期刊：
影响因子：
--
通讯作者：
Lässer C
中科院分区：
文献类型：
--
作者：
Karimi N;Cvjetkovic A;Jang SC;Crescitelli R;Hosseinpour Feizi MA;Nieuwland R;Lötvall J;Lässer C
从血液中分离细胞外小泡(EVS)对于了解循环EVS的生物学作用以及开发EVS作为疾病的生物标志物具有重要意义。然而，由于同时存在脂蛋白颗粒，血液是最难分离EV的体液之一。这项研究的目的是开发一种可靠的方法来从血液中分离和鉴定肠道病毒，并将血浆蛋白和脂蛋白颗粒的污染降至最低。采集健康受试者的血浆和血清，用尺寸排除层析(SEC)分离EVS，大部分颗粒存在于组分8-12，而大部分血浆蛋白存在于组分11-28。囊泡标记在组分7-11达到峰值；然而，相同的组分也含有脂蛋白颗粒。通过将密度垫与SEC相结合，进一步将脂蛋白颗粒与囊泡分离，从而提高了EVS的纯度，从而将脂蛋白颗粒的污染减少了100倍。利用这一新的分离方法，质谱仪共鉴定了血浆EVS中的1187个蛋白质，其中有几个蛋白质被称为EV相关蛋白，但在以往的血浆EVS蛋白质组学研究中尚未发现。这项研究表明，仅靠SEC无法将血浆EVS与脂蛋白颗粒完全分离。然而，SEC与密度垫相结合显著改善了EV与脂蛋白的分离，并允许对血浆EV的蛋白质组进行详细分析，从而使血液成为EV生物标记物发现的可行来源。本文的在线版本(10.1007/s000180182773-4)包含补充材料，可供授权用户使用。
The isolation of extracellular vesicles (EVs) from blood is of great importance to understand the biological role of circulating EVs and to develop EVs as biomarkers of disease. Due to the concurrent presence of lipoprotein particles, however, blood is one of the most difficult body fluids to isolate EVs from. The aim of this study was to develop a robust method to isolate and characterise EVs from blood with minimal contamination by plasma proteins and lipoprotein particles. Plasma and serum were collected from healthy subjects, and EVs were isolated by size-exclusion chromatography (SEC), with most particles being present in fractions 8–12, while the bulk of the plasma proteins was present in fractions 11–28. Vesicle markers peaked in fractions 7–11; however, the same fractions also contained lipoprotein particles. The purity of EVs was improved by combining a density cushion with SEC to further separate lipoprotein particles from the vesicles, which reduced the contamination of lipoprotein particles by 100-fold. Using this novel isolation procedure, a total of 1187 proteins were identified in plasma EVs by mass spectrometry, of which several proteins are known as EV-associated proteins but have hitherto not been identified in the previous proteomic studies of plasma EVs. This study shows that SEC alone is unable to completely separate plasma EVs from lipoprotein particles. However, combining SEC with a density cushion significantly improved the separation of EVs from lipoproteins and allowed for a detailed analysis of the proteome of plasma EVs, thus making blood a viable source for EV biomarker discovery. The online version of this article (10.1007/s00018-018-2773-4) contains supplementary material, which is available to authorized users.
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DOI：
10.1016/j.bbrc.2008.11.050
发表时间：
2009-01-16
影响因子：
3.1
作者：
Looze, Christopher;Yui, David;Leung, Lester;Ingham, Matthew;Kaler, Maryann;Yao, Xianglan;Wu, Wells W.;Shen, Rong-Fong;Daniels, Mathew P.;Levine, Stewart J.
通讯作者：
Levine, Stewart J.
影响因子：
8
作者：
Fischer, H;Polikarpov, I;Craievich, AF
通讯作者：
Craievich, AF
影响因子：
3.4
作者：
Kalra, Hina;Adda, Christopher G.;Mathivanan, Suresh
通讯作者：
Mathivanan, Suresh
影响因子：
21.3
作者：
Skog, Johan;Wuerdinger, Tom;van Rijn, Sjoerd;Meijer, Dimphna H.;Gainche, Laura;Sena-Esteves, Miguel;Curry, William T., Jr.;Carter, Bob S.;Krichevsky, Anna M.;Breakefield, Xandra O.
通讯作者：
Breakefield, Xandra O.
影响因子：
4.6
作者：
Sódar BW;Kittel Á;Pálóczi K;Vukman KV;Osteikoetxea X;Szabó-Taylor K;Németh A;Sperlágh B;Baranyai T;Giricz Z;Wiener Z;Turiák L;Drahos L;Pállinger É;Vékey K;Ferdinandy P;Falus A;Buzás EI
通讯作者：
Buzás EI