演題概要

ポスター発表

第1日 5月18日(水)  ポスター会場(月光)

細胞外小胞体“エクソソーム”の単離精製に関する比較検討

(住化分析セ)
o髙橋昭博丸谷曜子岡嶋孝太郎十亀祥久柳和則

Extracellular vesicles are spherical bi-layered nano-sized vesicles with 50-200 nm diameter released from all cell types, some molecular contents of which are a fingerprint of the releasing-cell types and of its physiological and oncogenic conditions. Hence, much attention has been paid to extracellular vesicles as diagnostic tools and for an ideal source in the discovery of new biomarkers. Recently, materials for isolating extracellular vesicles are commercially available as alternative to ultracentrifugation. The strategy and technique for isolating extracellular vesicles by these materials have based on physicochemical (size, density, and solubility) and biological (surface proteins and components of lipid membrane) properties of extracellular vesicles without requiring any expensive equipment. Our group performed a comprehensive evaluation of current methods for isolation of extracellular vesicles including ultracentrifugation, polymer-based precipitant, size exclusion column, immunoaffinity capture, and chelating agent. We employed a mass spectrometry-based proteomic approach to profile the extracellular vesicle-related proteins (ESCRT complex components, Rab GTPase, tetraspanins, trafficking, and cytoskeletal proteins), and a proteomic label-free emPAI approach that is directly proportional to protein contents in a sample. In this presentation, we will discuss characterizations of current methods for isolating extracellular vesicles on proteomic analysis as well as their advantages and disadvantages.