Evaluation of exosome separation from human serum by frit-inlet asymmetrical flow field-flow fractionation and multiangle light scattering

Analytica Chimica Acta
Young Beom KimMyeong Hee Moon

Abstract

Exosomes are extracellular vesicles that mediate intercellular communication, immune response, and tumour metastasis. However, exosome isolation from the blood is complicated because their size and density are similar to those of blood lipoproteins. Here, we employed field programming frit-inlet asymmetrical flow field-flow fractionation (FIAF4) coupled with multiangle light scattering (MALS) for the effective separation of exosomes from free unbound proteins and lipoproteins present in serum samples using different pre-treatment methods, namely, a commercial exosome isolation kit, ultracentrifugation (UC), and a simple centrifugation followed by ultrafiltration (UF). Sizes of the eluted exosomes, as calculated by MALS signals, approximated well with the results of batch dynamic light scattering of the collected fractions and with the sizes of polystyrene particles. Exosome separation from lipoproteins was validated by western blotting with several markers of exosomes and lipoproteins, followed by proteomic analysis using nanoflow ultrahigh-performance liquid chromatography-electrospray ionisation-tandem mass spectrometry. UC requires relatively large amounts of serum samples (at least 2 mL) but is more efficient at removing li...Continue Reading

References

Aug 3, 2002·Nature Reviews. Immunology·Clotilde ThérySebastian Amigorena
Oct 29, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Ilyong ParkMyeong Hee Moon
Feb 13, 2003·Journal of Chromatography. a·Pierluigi ReschiglianMyeong Hee Moon
Jan 30, 2008·Current Protocols in Cell Biology·Clotilde ThéryAled Clayton
Feb 17, 2009·Analytica Chimica Acta·Barbara RodaAldo Roda
Jun 5, 2012·Frontiers in Physiology·Fatemeh Momen-HeraviWinston Patrick Kuo
Feb 20, 2013·The Journal of Cell Biology·Graça Raposo, Willem Stoorvogel
Apr 8, 2014·Trends in Molecular Medicine·Pieter VaderMatthew J A Wood
Oct 4, 2014·Journal of Extracellular Vesicles·Anita N BöingRienk Nieuwland
May 26, 2015·Analytical Chemistry·Joon Seon YangMyeong Hee Moon
Aug 7, 2018·Journal of Proteome Research·Tingting Wang, Illarion V Turko

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Citations

Oct 8, 2020·Biomedical Chromatography : BMC·Edouard C Nice
Dec 15, 2020·Journal of Chromatography. a·Thanaporn LiangsupreeMarja-Liisa Riekkola
Jul 10, 2021·Frontiers in Cell and Developmental Biology·Yuxuan ZhongJun Fan
Aug 15, 2021·Biotechnology Advances·Saeedreza Zeibi Shirejini, Fatih Inci
Nov 2, 2021·ACS Nano·Sara Hassanpour TamrinArindom Sen
Nov 9, 2021·Mass Spectrometry Reviews·Iqbal JalaludinJeongkwon Kim
Nov 18, 2021·The Analyst·Takayuki KawaiKoji Otsuka

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