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Title: An LC-IMS-MS Platform Providing Increased Dynamic Range for High-Throughput Proteomic Studies

Abstract

A high-throughput approach and platform using 15 minute reversed-phase capillary liquid chromatography (RPLC) separations in conjunction with ion mobility spectrometry-mass spectrometry (IMS-MS) measurements was evaluated for the rapid analysis of complex proteomics samples. To test the separation quality of the short LC gradient, a sample was prepared by spiking twenty reference peptides at varying concentrations from 1 ng/mL to 10 µg/mL into a tryptic digest of mouse blood plasma and analyzed with both a LC-Linear Ion Trap Fourier Transform (FT) MS and LC-IMS-TOF MS. The LC-FT MS detected thirteen out of the twenty spiked peptides that had concentrations ≥100 ng/mL. In contrast, the drift time selected mass spectra from the LC-IMS-TOF MS analyses yielded identifications for nineteen of the twenty peptides with all spiking level present. The greater dynamic range of the LC-IMS-TOF MS system could be attributed to two factors. First, the LC-IMS-TOF MS system enabled drift time separation of the low concentration spiked peptides from the high concentration mouse peptide matrix components, reducing signal interference and background, and allowing species to be resolved that would otherwise be obscured by other components. Second, the automatic gain control (AGC) in the linear ion trap of the hybrid FT MS instrumentmore » limits the number of ions that are accumulated to reduce space charge effects, but in turn limits the achievable dynamic range compared to the TOF detector.« less

Authors:
; ; ; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
Sponsoring Org.:
USDOE
OSTI Identifier:
974911
Report Number(s):
PNNL-SA-67461
24698; 17797; 32707; 31590; KP1704020
DOE Contract Number:
AC05-76RL01830
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of Proteome Research, 9(2):997-1006
Country of Publication:
United States
Language:
English
Subject:
Ion mobility spectrometry; IMS-MS; LC-IMS-MS; high-throughput RPLC; ion-cyclotron resonance; capillary liquid-chromatography; TOF mass spectrometry; human plasma proteome; mobility spectrometry; biomarker discovery; RPLC separations; funnel trap; cancer; Environmental Molecular Sciences Laboratory

Citation Formats

Baker, Erin Shammel, Livesay, Eric A., Orton, Daniel J., Moore, Ronald J., Danielson, William F., Prior, David C., Ibrahim, Yehia M., Lamarche, Brian L., Mayampurath, Anoop M., Schepmoes, Athena A., Hopkins, Derek F., Tang, Keqi, Smith, Richard D., and Belov, Mikhail E. An LC-IMS-MS Platform Providing Increased Dynamic Range for High-Throughput Proteomic Studies. United States: N. p., 2010. Web. doi:10.1021/pr900888b.
Baker, Erin Shammel, Livesay, Eric A., Orton, Daniel J., Moore, Ronald J., Danielson, William F., Prior, David C., Ibrahim, Yehia M., Lamarche, Brian L., Mayampurath, Anoop M., Schepmoes, Athena A., Hopkins, Derek F., Tang, Keqi, Smith, Richard D., & Belov, Mikhail E. An LC-IMS-MS Platform Providing Increased Dynamic Range for High-Throughput Proteomic Studies. United States. doi:10.1021/pr900888b.
Baker, Erin Shammel, Livesay, Eric A., Orton, Daniel J., Moore, Ronald J., Danielson, William F., Prior, David C., Ibrahim, Yehia M., Lamarche, Brian L., Mayampurath, Anoop M., Schepmoes, Athena A., Hopkins, Derek F., Tang, Keqi, Smith, Richard D., and Belov, Mikhail E. Fri . "An LC-IMS-MS Platform Providing Increased Dynamic Range for High-Throughput Proteomic Studies". United States. doi:10.1021/pr900888b.
@article{osti_974911,
title = {An LC-IMS-MS Platform Providing Increased Dynamic Range for High-Throughput Proteomic Studies},
author = {Baker, Erin Shammel and Livesay, Eric A. and Orton, Daniel J. and Moore, Ronald J. and Danielson, William F. and Prior, David C. and Ibrahim, Yehia M. and Lamarche, Brian L. and Mayampurath, Anoop M. and Schepmoes, Athena A. and Hopkins, Derek F. and Tang, Keqi and Smith, Richard D. and Belov, Mikhail E.},
abstractNote = {A high-throughput approach and platform using 15 minute reversed-phase capillary liquid chromatography (RPLC) separations in conjunction with ion mobility spectrometry-mass spectrometry (IMS-MS) measurements was evaluated for the rapid analysis of complex proteomics samples. To test the separation quality of the short LC gradient, a sample was prepared by spiking twenty reference peptides at varying concentrations from 1 ng/mL to 10 µg/mL into a tryptic digest of mouse blood plasma and analyzed with both a LC-Linear Ion Trap Fourier Transform (FT) MS and LC-IMS-TOF MS. The LC-FT MS detected thirteen out of the twenty spiked peptides that had concentrations ≥100 ng/mL. In contrast, the drift time selected mass spectra from the LC-IMS-TOF MS analyses yielded identifications for nineteen of the twenty peptides with all spiking level present. The greater dynamic range of the LC-IMS-TOF MS system could be attributed to two factors. First, the LC-IMS-TOF MS system enabled drift time separation of the low concentration spiked peptides from the high concentration mouse peptide matrix components, reducing signal interference and background, and allowing species to be resolved that would otherwise be obscured by other components. Second, the automatic gain control (AGC) in the linear ion trap of the hybrid FT MS instrument limits the number of ions that are accumulated to reduce space charge effects, but in turn limits the achievable dynamic range compared to the TOF detector.},
doi = {10.1021/pr900888b},
journal = {Journal of Proteome Research, 9(2):997-1006},
number = ,
volume = ,
place = {United States},
year = {Fri Feb 05 00:00:00 EST 2010},
month = {Fri Feb 05 00:00:00 EST 2010}
}