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 - Yeul, Hong Kim
AU - Koo, Kyung Hee
AU - Ho, Zoon Chae
N1 - Funding Information:
This work was supported in part by a Cancer Center Support Grant (CA016056-27) awarded to RPCI from the National Cancer Institute. It was also supported by grants from the National Cancer Institute (CA109480), Human Proteome Organization, and Korean Ministry of Science and Welfare.
PY - 2005
Y1 - 2005
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
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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 -