Stable, reproducible, and automated capillary zone electrophoresis-tandem mass spectrometry system with an electrokinetically pumped sheath-flow nanospray interface.
A PrinCE autosampler was coupled to a Q-Exactive mass spectrometer by an electrokinetically pumped sheath-flow nanospray interface to perform automated capillary zone electrophoresis-electrospray ionization-tandem mass spectrometry (CZE-ESI-MS/MS). 20ng aliquots of an Escherichia coli digest were injected to evaluate the system. Eight sequential injections over an 8-h period identified 1115±70 (relative standard deviation, RSD=6%) peptides and 270±8 (RSD=3%) proteins per run. The average RSDs of migration time, peak intensity, and peak area were 3%, 24% and 19%, respectively, for 340 peptides with high intensity. This is the first report of an automated CZE-ESI-MS/MS system using the electrokinetically pumped sheath-flow nanospray interface. The results demonstrate that this system is capable of reproducibly identifying over 1000 peptides from an E. coli tryptic digest in a 1-h analysis time.
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA.
Guijie
G
Zhu
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA.
Liangliang
L
Sun
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA.
Xiaojing
X
Yan
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA. Electronic address: ndovichi@nd.edu.
Norman J
NJ
Dovichi
D001331:Automation; D019075:Electrophoresis, Capillary; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D036103:Nanotechnology / Q000295:instrumentation*; D010455:Peptides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism
D029968:Escherichia coli Proteins; D010455:Peptides; D014357:Trypsin
Capillary zone electrophoresis; Proteomics