16 research outputs found
Quantitative Multiple Reaction Monitoring of Peptide Abundance Introduced via a Capillary Zone Electrophoresis–Electrospray Interface
We demonstrate the use of capillary zone electrophoresis
with an
electrokinetic sheath-flow electrospray interface coupled to a triple-quadrupole
mass spectrometer for the accurate and precise quantification of Leu-enkephalin
in a complex mixture using multiple-reaction monitoring (MRM). Assay
time is <6 min, with no re-equilibration required between runs.
A standard curve of Leu-enkephalin was performed in the presence of
a background tryptic digest of bovine albumin. We demonstrate reasonably
reproducible peak heights (21% relative standard deviation), retention
times (better than 1% relative standard deviation), and robust electrospray
quality. Our limit of detection (3σ) was 60 pM, which corresponds
to the injection of 335 zmol of peptide. This is a 10–20-fold
improvement in mass sensitivity than we have obtained by nano HPLC/MRM
and substantially better than reported for LC/MS/MS. Further quantification
was performed in the presence of stable-isotope-labeled versions of
the peptides; under these conditions, linearity was observed across
nearly 4 orders of magnitude. The concentration detection limit was
240 pM for the stable-isotope-labeled quantification
Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry as an Alternative Proteomics Platform to Ultraperformance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry for Samples of Intermediate Complexity
We demonstrate the use of capillary zone electrophoresis
with an
electrokinetically pumped sheath-flow electrospray interface for the
analysis of a tryptic digest of a sample of intermediate protein complexity,
the secreted protein fraction of <i>Mycobacterium marinum</i>. For electrophoretic analysis, 11 fractions were generated from
the sample using reverse-phase liquid chromatography; each fraction
was analyzed by CZE-ESI-MS/MS, and 334 peptides corresponding to 140
proteins were identified in 165 min of mass spectrometer time at 95%
confidence (FDR < 0.15%). In comparison, 388 peptides corresponding
to 134 proteins were identified in 180 min of mass spectrometer time
by triplicate UPLC-ESI-MS/MS analyses, each using 250 ng of the unfractionated
peptide mixture, at 95% confidence (FDR < 0.15%). Overall, 62%
of peptides identified in CZE-ESI-MS/MS and 67% in UPLC-ESI-MS/MS
were unique. CZE-ESI-MS/MS favored basic and hydrophilic peptides
with low molecular masses. Combining the two data sets increased the
number of unique peptides by 53%. Our approach identified more than
twice as many proteins as the previous record for capillary electrophoresis
proteome analysis. CE-ESI-MS/MS is a useful tool for the analysis
of proteome samples of intermediate complexity
Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry as an Alternative Proteomics Platform to Ultraperformance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry for Samples of Intermediate Complexity
We demonstrate the use of capillary zone electrophoresis
with an
electrokinetically pumped sheath-flow electrospray interface for the
analysis of a tryptic digest of a sample of intermediate protein complexity,
the secreted protein fraction of <i>Mycobacterium marinum</i>. For electrophoretic analysis, 11 fractions were generated from
the sample using reverse-phase liquid chromatography; each fraction
was analyzed by CZE-ESI-MS/MS, and 334 peptides corresponding to 140
proteins were identified in 165 min of mass spectrometer time at 95%
confidence (FDR < 0.15%). In comparison, 388 peptides corresponding
to 134 proteins were identified in 180 min of mass spectrometer time
by triplicate UPLC-ESI-MS/MS analyses, each using 250 ng of the unfractionated
peptide mixture, at 95% confidence (FDR < 0.15%). Overall, 62%
of peptides identified in CZE-ESI-MS/MS and 67% in UPLC-ESI-MS/MS
were unique. CZE-ESI-MS/MS favored basic and hydrophilic peptides
with low molecular masses. Combining the two data sets increased the
number of unique peptides by 53%. Our approach identified more than
twice as many proteins as the previous record for capillary electrophoresis
proteome analysis. CE-ESI-MS/MS is a useful tool for the analysis
of proteome samples of intermediate complexity
Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry as an Alternative Proteomics Platform to Ultraperformance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry for Samples of Intermediate Complexity
We demonstrate the use of capillary zone electrophoresis
with an
electrokinetically pumped sheath-flow electrospray interface for the
analysis of a tryptic digest of a sample of intermediate protein complexity,
the secreted protein fraction of <i>Mycobacterium marinum</i>. For electrophoretic analysis, 11 fractions were generated from
the sample using reverse-phase liquid chromatography; each fraction
was analyzed by CZE-ESI-MS/MS, and 334 peptides corresponding to 140
proteins were identified in 165 min of mass spectrometer time at 95%
confidence (FDR < 0.15%). In comparison, 388 peptides corresponding
to 134 proteins were identified in 180 min of mass spectrometer time
by triplicate UPLC-ESI-MS/MS analyses, each using 250 ng of the unfractionated
peptide mixture, at 95% confidence (FDR < 0.15%). Overall, 62%
of peptides identified in CZE-ESI-MS/MS and 67% in UPLC-ESI-MS/MS
were unique. CZE-ESI-MS/MS favored basic and hydrophilic peptides
with low molecular masses. Combining the two data sets increased the
number of unique peptides by 53%. Our approach identified more than
twice as many proteins as the previous record for capillary electrophoresis
proteome analysis. CE-ESI-MS/MS is a useful tool for the analysis
of proteome samples of intermediate complexity
Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry as an Alternative Proteomics Platform to Ultraperformance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry for Samples of Intermediate Complexity
We demonstrate the use of capillary zone electrophoresis
with an
electrokinetically pumped sheath-flow electrospray interface for the
analysis of a tryptic digest of a sample of intermediate protein complexity,
the secreted protein fraction of <i>Mycobacterium marinum</i>. For electrophoretic analysis, 11 fractions were generated from
the sample using reverse-phase liquid chromatography; each fraction
was analyzed by CZE-ESI-MS/MS, and 334 peptides corresponding to 140
proteins were identified in 165 min of mass spectrometer time at 95%
confidence (FDR < 0.15%). In comparison, 388 peptides corresponding
to 134 proteins were identified in 180 min of mass spectrometer time
by triplicate UPLC-ESI-MS/MS analyses, each using 250 ng of the unfractionated
peptide mixture, at 95% confidence (FDR < 0.15%). Overall, 62%
of peptides identified in CZE-ESI-MS/MS and 67% in UPLC-ESI-MS/MS
were unique. CZE-ESI-MS/MS favored basic and hydrophilic peptides
with low molecular masses. Combining the two data sets increased the
number of unique peptides by 53%. Our approach identified more than
twice as many proteins as the previous record for capillary electrophoresis
proteome analysis. CE-ESI-MS/MS is a useful tool for the analysis
of proteome samples of intermediate complexity
Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry as an Alternative Proteomics Platform to Ultraperformance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry for Samples of Intermediate Complexity
We demonstrate the use of capillary zone electrophoresis
with an
electrokinetically pumped sheath-flow electrospray interface for the
analysis of a tryptic digest of a sample of intermediate protein complexity,
the secreted protein fraction of <i>Mycobacterium marinum</i>. For electrophoretic analysis, 11 fractions were generated from
the sample using reverse-phase liquid chromatography; each fraction
was analyzed by CZE-ESI-MS/MS, and 334 peptides corresponding to 140
proteins were identified in 165 min of mass spectrometer time at 95%
confidence (FDR < 0.15%). In comparison, 388 peptides corresponding
to 134 proteins were identified in 180 min of mass spectrometer time
by triplicate UPLC-ESI-MS/MS analyses, each using 250 ng of the unfractionated
peptide mixture, at 95% confidence (FDR < 0.15%). Overall, 62%
of peptides identified in CZE-ESI-MS/MS and 67% in UPLC-ESI-MS/MS
were unique. CZE-ESI-MS/MS favored basic and hydrophilic peptides
with low molecular masses. Combining the two data sets increased the
number of unique peptides by 53%. Our approach identified more than
twice as many proteins as the previous record for capillary electrophoresis
proteome analysis. CE-ESI-MS/MS is a useful tool for the analysis
of proteome samples of intermediate complexity
Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry as an Alternative Proteomics Platform to Ultraperformance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry for Samples of Intermediate Complexity
We demonstrate the use of capillary zone electrophoresis
with an
electrokinetically pumped sheath-flow electrospray interface for the
analysis of a tryptic digest of a sample of intermediate protein complexity,
the secreted protein fraction of <i>Mycobacterium marinum</i>. For electrophoretic analysis, 11 fractions were generated from
the sample using reverse-phase liquid chromatography; each fraction
was analyzed by CZE-ESI-MS/MS, and 334 peptides corresponding to 140
proteins were identified in 165 min of mass spectrometer time at 95%
confidence (FDR < 0.15%). In comparison, 388 peptides corresponding
to 134 proteins were identified in 180 min of mass spectrometer time
by triplicate UPLC-ESI-MS/MS analyses, each using 250 ng of the unfractionated
peptide mixture, at 95% confidence (FDR < 0.15%). Overall, 62%
of peptides identified in CZE-ESI-MS/MS and 67% in UPLC-ESI-MS/MS
were unique. CZE-ESI-MS/MS favored basic and hydrophilic peptides
with low molecular masses. Combining the two data sets increased the
number of unique peptides by 53%. Our approach identified more than
twice as many proteins as the previous record for capillary electrophoresis
proteome analysis. CE-ESI-MS/MS is a useful tool for the analysis
of proteome samples of intermediate complexity
Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry as an Alternative Proteomics Platform to Ultraperformance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry for Samples of Intermediate Complexity
We demonstrate the use of capillary zone electrophoresis
with an
electrokinetically pumped sheath-flow electrospray interface for the
analysis of a tryptic digest of a sample of intermediate protein complexity,
the secreted protein fraction of <i>Mycobacterium marinum</i>. For electrophoretic analysis, 11 fractions were generated from
the sample using reverse-phase liquid chromatography; each fraction
was analyzed by CZE-ESI-MS/MS, and 334 peptides corresponding to 140
proteins were identified in 165 min of mass spectrometer time at 95%
confidence (FDR < 0.15%). In comparison, 388 peptides corresponding
to 134 proteins were identified in 180 min of mass spectrometer time
by triplicate UPLC-ESI-MS/MS analyses, each using 250 ng of the unfractionated
peptide mixture, at 95% confidence (FDR < 0.15%). Overall, 62%
of peptides identified in CZE-ESI-MS/MS and 67% in UPLC-ESI-MS/MS
were unique. CZE-ESI-MS/MS favored basic and hydrophilic peptides
with low molecular masses. Combining the two data sets increased the
number of unique peptides by 53%. Our approach identified more than
twice as many proteins as the previous record for capillary electrophoresis
proteome analysis. CE-ESI-MS/MS is a useful tool for the analysis
of proteome samples of intermediate complexity
Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry as an Alternative Proteomics Platform to Ultraperformance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry for Samples of Intermediate Complexity
We demonstrate the use of capillary zone electrophoresis
with an
electrokinetically pumped sheath-flow electrospray interface for the
analysis of a tryptic digest of a sample of intermediate protein complexity,
the secreted protein fraction of <i>Mycobacterium marinum</i>. For electrophoretic analysis, 11 fractions were generated from
the sample using reverse-phase liquid chromatography; each fraction
was analyzed by CZE-ESI-MS/MS, and 334 peptides corresponding to 140
proteins were identified in 165 min of mass spectrometer time at 95%
confidence (FDR < 0.15%). In comparison, 388 peptides corresponding
to 134 proteins were identified in 180 min of mass spectrometer time
by triplicate UPLC-ESI-MS/MS analyses, each using 250 ng of the unfractionated
peptide mixture, at 95% confidence (FDR < 0.15%). Overall, 62%
of peptides identified in CZE-ESI-MS/MS and 67% in UPLC-ESI-MS/MS
were unique. CZE-ESI-MS/MS favored basic and hydrophilic peptides
with low molecular masses. Combining the two data sets increased the
number of unique peptides by 53%. Our approach identified more than
twice as many proteins as the previous record for capillary electrophoresis
proteome analysis. CE-ESI-MS/MS is a useful tool for the analysis
of proteome samples of intermediate complexity
Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry as an Alternative Proteomics Platform to Ultraperformance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry for Samples of Intermediate Complexity
We demonstrate the use of capillary zone electrophoresis
with an
electrokinetically pumped sheath-flow electrospray interface for the
analysis of a tryptic digest of a sample of intermediate protein complexity,
the secreted protein fraction of <i>Mycobacterium marinum</i>. For electrophoretic analysis, 11 fractions were generated from
the sample using reverse-phase liquid chromatography; each fraction
was analyzed by CZE-ESI-MS/MS, and 334 peptides corresponding to 140
proteins were identified in 165 min of mass spectrometer time at 95%
confidence (FDR < 0.15%). In comparison, 388 peptides corresponding
to 134 proteins were identified in 180 min of mass spectrometer time
by triplicate UPLC-ESI-MS/MS analyses, each using 250 ng of the unfractionated
peptide mixture, at 95% confidence (FDR < 0.15%). Overall, 62%
of peptides identified in CZE-ESI-MS/MS and 67% in UPLC-ESI-MS/MS
were unique. CZE-ESI-MS/MS favored basic and hydrophilic peptides
with low molecular masses. Combining the two data sets increased the
number of unique peptides by 53%. Our approach identified more than
twice as many proteins as the previous record for capillary electrophoresis
proteome analysis. CE-ESI-MS/MS is a useful tool for the analysis
of proteome samples of intermediate complexity