Proteomic analysis of exosomes from human neural stem cells by flow field-flow fractionation and nanoflow liquid chromatography-tandem mass spectrometry

Dukjin Kang, Sunok Oh, Sung Min Ahn, Bong Hee Lee, Myeong Hee Moon

Research output: Contribution to journalArticle

90 Citations (Scopus)

Abstract

Exosomes, small membrane vesicles secreted by a multitude of cell types, are involved in a wide range of physiological roles such as intercellular communication, membrane exchange between cells, and degradation as an alternative to lysosomes. Because of the small size of exosomes (30-100 nm) and the limitations of common separation procedures including ultracentrifugation and flow cytometry, size-based fractionation of exosomes has been challenging. In this study, we used flow field-flow fractionation (FIFFF) to fractionate exosomes according to differences in hydrodynamic diameter. The exosome fractions collected from FIFFF runs were examined by transmission electron microscopy (TEM) to morphologically confirm their identification as exosomes. Exosomal lysates of each fraction were digested and analyzed using nanoflow LC-ESI-MS-MS for protein identification. FIFFF, coupled with mass spectrometry, allows nanoscale size-based fractionation of exosomes and is more applicable to primary cells and stem cells since it requires much less starting material than conventional gel-based separation, in-gel digestion and the MS-MS method.

Original languageEnglish
Pages (from-to)3475-3480
Number of pages6
JournalJournal of Proteome Research
Volume7
Issue number8
DOIs
Publication statusPublished - 2008 Aug 1

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Field Flow Fractionation
Exosomes
Neural Stem Cells
Liquid chromatography
Fractionation
Tandem Mass Spectrometry
Stem cells
Liquid Chromatography
Proteomics
Mass spectrometry
Flow fields
Gels
Membranes
Flow cytometry
Ion exchange
Hydrodynamics
Transmission electron microscopy
Degradation
Communication
Ultracentrifugation

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Chemistry(all)

Cite this

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abstract = "Exosomes, small membrane vesicles secreted by a multitude of cell types, are involved in a wide range of physiological roles such as intercellular communication, membrane exchange between cells, and degradation as an alternative to lysosomes. Because of the small size of exosomes (30-100 nm) and the limitations of common separation procedures including ultracentrifugation and flow cytometry, size-based fractionation of exosomes has been challenging. In this study, we used flow field-flow fractionation (FIFFF) to fractionate exosomes according to differences in hydrodynamic diameter. The exosome fractions collected from FIFFF runs were examined by transmission electron microscopy (TEM) to morphologically confirm their identification as exosomes. Exosomal lysates of each fraction were digested and analyzed using nanoflow LC-ESI-MS-MS for protein identification. FIFFF, coupled with mass spectrometry, allows nanoscale size-based fractionation of exosomes and is more applicable to primary cells and stem cells since it requires much less starting material than conventional gel-based separation, in-gel digestion and the MS-MS method.",
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Proteomic analysis of exosomes from human neural stem cells by flow field-flow fractionation and nanoflow liquid chromatography-tandem mass spectrometry. / Kang, Dukjin; Oh, Sunok; Ahn, Sung Min; Lee, Bong Hee; Moon, Myeong Hee.

In: Journal of Proteome Research, Vol. 7, No. 8, 01.08.2008, p. 3475-3480.

Research output: Contribution to journalArticle

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AU - Moon, Myeong Hee

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