Abstract
We describe the use of a single pump capillary-LC system (flow rate: 1-100 μL/min) to perform both capillary-LC-μESI-MS/MS and nano-LC-nano ESI-MS/MS (flow rate: 50-1000 nL/min) analysis and its applications in proteomics. A highly constant nanoflow (180 nL/min) delivery has been achieved by using a capillary binary-pump with electronic flow control and a flow splitter with compensation for the viscosity changes during gradient elution. A column switching technique was successfully used in nano-LC-nanoESI-MS/MS analysis to provide two flow paths, one for sample loading with a flow rate of 15 μL/min directly from the capillary pump, the other for nanoflow gradient elution with a flow rate of 180 nL/min after flow splitting. In comparison with the capillary-LC-μESI-MS/MS at a flow rate of 3 μL/min, this nano-LC-nano ESI-MS/MS analysis improved sensitivity by more than 50 times. It has been successfully used to identify protein spots from 2DGE, as well as protein bands from 1DGE, after in-gel tryptic digestion. Similar to any commercial nano-LC system, this novel nano-LC configuration can be used to perform MudPIT analysis, as well as offline capillary-SCX/nano-RPLC-nanoESI-MS/MS analysis. Our data has shown the unnecessary need for either a second pump or an expensive nano-pump to do nano-LC analysis as most commercial nano-LC system requires, and the increased sensitivity, functionality, and flexibility of a simple capillary-LC system, which is very useful to typical analytical laboratories for multiple-purpose usages of limited resources.
| Original language | English |
|---|---|
| Pages (from-to) | 1271-1289 |
| Number of pages | 19 |
| Journal | Journal of Liquid Chromatography and Related Technologies |
| Volume | 28 |
| Issue number | 9 |
| DOIs | |
| Publication status | Published - 2005 May 23 |
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Keywords
- Capillary-LC-μESI-MS/MS
- Column switching
- Human plasma
- Nano-LC-nano ESI-MS/MS
- Proteomics
- Two-dimensional liquid chromatography
ASJC Scopus subject areas
- Analytical Chemistry
- Clinical Biochemistry
Cite this
Capillary-LC-μESI-MS/MS and nano-LC-nano ESI-MS/MS analysis using a single binary pump capillary LC system : Applications in proteomics. / Song, Liguo; Choi, Kyoung Soo; Park, Young Mee; Kazim, A. Latif; Marlar, Khin; Kong, Eung Sik; Park, Eun Mi; Kim, Yeul Hong; Koo, Kyung Hee; Ho, Zoon Chae.
In: Journal of Liquid Chromatography and Related Technologies, Vol. 28, No. 9, 23.05.2005, p. 1271-1289.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Capillary-LC-μESI-MS/MS and nano-LC-nano ESI-MS/MS analysis using a single binary pump capillary LC system
T2 - Applications in proteomics
AU - Song, Liguo
AU - Choi, Kyoung Soo
AU - Park, Young Mee
AU - Kazim, A. Latif
AU - Marlar, Khin
AU - Kong, Eung Sik
AU - Park, Eun Mi
AU - Kim, Yeul Hong
AU - Koo, Kyung Hee
AU - Ho, Zoon Chae
PY - 2005/5/23
Y1 - 2005/5/23
N2 - We describe the use of a single pump capillary-LC system (flow rate: 1-100 μL/min) to perform both capillary-LC-μESI-MS/MS and nano-LC-nano ESI-MS/MS (flow rate: 50-1000 nL/min) analysis and its applications in proteomics. A highly constant nanoflow (180 nL/min) delivery has been achieved by using a capillary binary-pump with electronic flow control and a flow splitter with compensation for the viscosity changes during gradient elution. A column switching technique was successfully used in nano-LC-nanoESI-MS/MS analysis to provide two flow paths, one for sample loading with a flow rate of 15 μL/min directly from the capillary pump, the other for nanoflow gradient elution with a flow rate of 180 nL/min after flow splitting. In comparison with the capillary-LC-μESI-MS/MS at a flow rate of 3 μL/min, this nano-LC-nano ESI-MS/MS analysis improved sensitivity by more than 50 times. It has been successfully used to identify protein spots from 2DGE, as well as protein bands from 1DGE, after in-gel tryptic digestion. Similar to any commercial nano-LC system, this novel nano-LC configuration can be used to perform MudPIT analysis, as well as offline capillary-SCX/nano-RPLC-nanoESI-MS/MS analysis. Our data has shown the unnecessary need for either a second pump or an expensive nano-pump to do nano-LC analysis as most commercial nano-LC system requires, and the increased sensitivity, functionality, and flexibility of a simple capillary-LC system, which is very useful to typical analytical laboratories for multiple-purpose usages of limited resources.
AB - We describe the use of a single pump capillary-LC system (flow rate: 1-100 μL/min) to perform both capillary-LC-μESI-MS/MS and nano-LC-nano ESI-MS/MS (flow rate: 50-1000 nL/min) analysis and its applications in proteomics. A highly constant nanoflow (180 nL/min) delivery has been achieved by using a capillary binary-pump with electronic flow control and a flow splitter with compensation for the viscosity changes during gradient elution. A column switching technique was successfully used in nano-LC-nanoESI-MS/MS analysis to provide two flow paths, one for sample loading with a flow rate of 15 μL/min directly from the capillary pump, the other for nanoflow gradient elution with a flow rate of 180 nL/min after flow splitting. In comparison with the capillary-LC-μESI-MS/MS at a flow rate of 3 μL/min, this nano-LC-nano ESI-MS/MS analysis improved sensitivity by more than 50 times. It has been successfully used to identify protein spots from 2DGE, as well as protein bands from 1DGE, after in-gel tryptic digestion. Similar to any commercial nano-LC system, this novel nano-LC configuration can be used to perform MudPIT analysis, as well as offline capillary-SCX/nano-RPLC-nanoESI-MS/MS analysis. Our data has shown the unnecessary need for either a second pump or an expensive nano-pump to do nano-LC analysis as most commercial nano-LC system requires, and the increased sensitivity, functionality, and flexibility of a simple capillary-LC system, which is very useful to typical analytical laboratories for multiple-purpose usages of limited resources.
KW - Capillary-LC-μESI-MS/MS
KW - Column switching
KW - Human plasma
KW - Nano-LC-nano ESI-MS/MS
KW - Proteomics
KW - Two-dimensional liquid chromatography
UR - http://www.scopus.com/inward/record.url?scp=20844431557&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=20844431557&partnerID=8YFLogxK
U2 - 10.1081/JLC-200054754
DO - 10.1081/JLC-200054754
M3 - Article
AN - SCOPUS:20844431557
VL - 28
SP - 1271
EP - 1289
JO - Journal of Liquid Chromatography and Related Technologies
JF - Journal of Liquid Chromatography and Related Technologies
SN - 1082-6076
IS - 9
ER -