Rapid isolation of extracellular vesicles from cell culture and biological fluids using a synthetic peptide with specific affinity for heat shock proteins.
Rapid isolation of extracellular vesicles from cell culture and biological fluids using a synthetic peptide with specific affinity for heat shock proteins.
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DOI：
10.1371/journal.pone.0110443
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Ouellette RJ
中科院分区：
文献类型：
--
作者：
Ghosh A;Davey M;Chute IC;Griffiths SG;Lewis S;Chacko S;Barnett D;Crapoulet N;Fournier S;Joy A;Caissie MC;Ferguson AD;Daigle M;Meli MV;Lewis SM;Ouellette RJ
最近的研究表明，细胞外囊泡是许多临床应用的重要来源材料，包括微创疾病诊断。然而，快速和简单的细胞外囊泡收集的挑战阻碍了它们的应用。我们已经开发并验证了一类新的肽（我们命名为venceremin或Vn），其对典型的热休克蛋白表现出不依赖于核苷酸的特异性亲和力。通过电子显微镜、原子力显微镜、核酸货物测序、蛋白质组学分析、免疫印迹和纳米颗粒跟踪分析，验证了Vn肽特异性且有效地从细胞培养生长培养基、血浆和尿液中捕获含HSP的细胞外囊泡。所有这些分析都证实了由Vn肽捕获的物质与通过标准超离心方法纯化的物质相当。我们表明，VN肽是一个有用的工具，使用标准的实验室设备的细胞外囊泡的快速分离。此外，Vn肽适用于不同的平台，因此代表了针对研究和临床应用的细胞外囊泡分离挑战的优秀解决方案。
Recent studies indicate that extracellular vesicles are an important source material for many clinical applications, including minimally-invasive disease diagnosis. However, challenges for rapid and simple extracellular vesicle collection have hindered their application. We have developed and validated a novel class of peptides (which we named venceremin, or Vn) that exhibit nucleotide-independent specific affinity for canonical heat shock proteins. The Vn peptides were validated to specifically and efficiently capture HSP-containing extracellular vesicles from cell culture growth media, plasma, and urine by electron microscopy, atomic force microscopy, sequencing of nucleic acid cargo, proteomic profiling, immunoblotting, and nanoparticle tracking analysis. All of these analyses confirmed the material captured by the Vn peptides was comparable to those purified by the standard ultracentrifugation method. We show that the Vn peptides are a useful tool for the rapid isolation of extracellular vesicles using standard laboratory equipment. Moreover, the Vn peptides are adaptable to diverse platforms and therefore represent an excellent solution to the challenge of extracellular vesicle isolation for research and clinical applications.
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DOI：
10.1111/j.1600-0854.2009.01006.x
发表时间：
2010-01
期刊：
Traffic (Copenhagen, Denmark)
影响因子：
--
作者：
Lenassi M;Cagney G;Liao M;Vaupotic T;Bartholomeeusen K;Cheng Y;Krogan NJ;Plemenitas A;Peterlin BM
通讯作者：
Peterlin BM
影响因子：
3.8
作者：
De Maio, Antonio
通讯作者：
De Maio, Antonio
DOI：
10.1073/pnas.0308413101
发表时间：
2004-06-29
影响因子：
11.1
作者：
Fevrier, B;Vilette, D;Raposo, G
通讯作者：
Raposo, G
DOI：
10.1165/rcmb.2003-0238oc
发表时间：
2004-07-01
影响因子：
6.4
作者：
Bard, MP;Hegmans, JP;Lambrecht, BN
通讯作者：
Lambrecht, BN
影响因子：
4.8
作者：
Lancaster, GI;Febbraio, MA
通讯作者：
Febbraio, MA