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 language | English |
|---|---|
| Pages (from-to) | 3475-3480 |
| Number of pages | 6 |
| Journal | Journal of Proteome Research |
| Volume | 7 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - 2008 Aug 1 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Chemistry(all)
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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 journal › Article
TY - JOUR
T1 - Proteomic analysis of exosomes from human neural stem cells by flow field-flow fractionation and nanoflow liquid chromatography-tandem mass spectrometry
AU - Kang, Dukjin
AU - Oh, Sunok
AU - Ahn, Sung Min
AU - Lee, Bong Hee
AU - Moon, Myeong Hee
PY - 2008/8/1
Y1 - 2008/8/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=53049107224&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=53049107224&partnerID=8YFLogxK
U2 - 10.1021/pr800225z
DO - 10.1021/pr800225z
M3 - Article
C2 - 18570454
AN - SCOPUS:53049107224
VL - 7
SP - 3475
EP - 3480
JO - Journal of Proteome Research
JF - Journal of Proteome Research
SN - 1535-3893
IS - 8
ER -