用于人类和鼠类细胞外囊泡和颗粒的跨平台特征描述的方案。

Protocol for cross-platform characterization of human and murine extracellular vesicles and particles.

机构信息

Children's Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children's Health, Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA; Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden.

Children's Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children's Health, Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA; Yonsei Cancer Center, Division of Medical Oncology, Department of Internal Medicine, Graduate School of Medical Science, Brain Korea 21 FOUR Project, Yonsei University College of Medicine, Seoul, Korea.

出版信息

STAR Protoc. 2024 Mar 15;5(1):102754. doi: 10.1016/j.xpro.2023.102754. Epub 2023 Dec 13.

DOI:10.1016/j.xpro.2023.102754

PMID:38096060

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10762520/

Abstract

Characterization of isolated extracellular vesicles and particles (EVPs) is crucial for determining functions and biomarker potential. Here, we present a protocol to analyze size, number, morphology, and EVP protein cargo and to validate EVP proteins in both humans and mice. We describe steps for nanoparticle tracking analysis, transmission electron microscopy, single-EVP immunodetection, EVP proteomic mass spectrometry and bioinformatic analysis, and EVP protein validation by ExoELISA and western blot analysis. This allows for EVP cross-validation across different platforms. For complete details on the use and execution of this protocol, please refer to Hoshino et al..

摘要

分离的细胞外囊泡和颗粒（EVPs）的特征对于确定其功能和生物标志物潜力至关重要。在这里，我们提供了一个分析大小、数量、形态和 EVPs 蛋白负荷的方案，并在人类和小鼠中验证了 EVPs 蛋白。我们描述了纳米颗粒跟踪分析、透射电子显微镜、单个 EVPs 免疫检测、EVPs 蛋白质组学质谱和生物信息学分析的步骤，以及通过 ExoELISA 和 Western blot 分析验证 EVPs 蛋白的步骤。这允许在不同平台上进行 EVPs 的交叉验证。有关此方案使用和执行的完整详细信息，请参阅 Hoshino 等人的研究。