Rapid Isolation of Extracellular Vesicles Using a Hydrophilic Porous Silica Gel-Based Size-Exclusion Chromatography Column

Jun Yoshitake, Mayuko Azami, Haruka Sei, Daisuke Onoshima, Kumiko Takahashi, Akiyoshi Hirayama, Koji Uchida, Yoshinobu Baba, Takahiro Shibata

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

Extracellular vesicles (EVs) are nanoscale lipid bilayer vesicles released by almost all cell types and can be found in biological fluids, such as blood and urine. EVs play an important role in various physiological and pathological processes via cell-cell communication, highlighting their potential applications as diagnostic markers for diseases and therapeutic drug delivery carriers. Although various methods have been developed for the isolation of EVs from biological fluids, most of them exhibit major limitations, including low purity, long processing times, and high cost. In this study, we developed a size-exclusion chromatography (SEC) column device using hydrophilic porous silica gel (PSG). Owing to the resistance to pressure of the device, a rapid system for EV isolation was developed by connecting it to a flash liquid chromatography system furnished with a UV detector and a fraction collector. This system can be used for the real-time monitoring of eluted EVs by UV absorption without further analysis and separation of high-purity EVs from urine samples with high durability, reusability, and reproducibility. In addition, there were no significant differences between the PSG column- and conventional SEC column-isolated EVs in the proteome profiles and cellular uptake activities, suggesting the good quality of the EVs isolated by the PSG column. These findings suggest that the PSG column device offers an effective and rapid method for the isolation of intact EVs from biological fluids.

Original languageEnglish
Pages (from-to)13676-13681
Number of pages6
JournalAnalytical Chemistry
Volume94
Issue number40
DOIs
Publication statusPublished - 2022 Oct 11

ASJC Scopus subject areas

  • Analytical Chemistry

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