Research ExplorerPublicationsDisplacement chromatography as first separating step in onli...

Displacement chromatography as first separating step in online two-dimensional liquid chromatography coupled to mass spectrometry analysis of a complex protein sample--the proteome of neutrophils

Standard

Displacement chromatography as first separating step in online two-dimensional liquid chromatography coupled to mass spectrometry analysis of a complex protein sample--the proteome of neutrophils. / Trusch, Maria; Tillack, Kati; Kwiatkowski, Marcel; Bertsch, Andreas; Ahrends, Robert; Kohlbacher, Oliver; Martin, Roland; Sospedra, Mireia; Schlüter, Hartmut.

In: J CHROMATOGR A, Vol. 1232, 06.04.2012, p. 288-294.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Trusch, M, Tillack, K, Kwiatkowski, M, Bertsch, A, Ahrends, R, Kohlbacher, O, Martin, R, Sospedra, M & Schlüter, H 2012, 'Displacement chromatography as first separating step in online two-dimensional liquid chromatography coupled to mass spectrometry analysis of a complex protein sample--the proteome of neutrophils', J CHROMATOGR A, vol. 1232, pp. 288-294. https://doi.org/10.1016/j.chroma.2012.02.029

APA

Trusch, M., Tillack, K., Kwiatkowski, M., Bertsch, A., Ahrends, R., Kohlbacher, O., Martin, R., Sospedra, M., & Schlüter, H. (2012). Displacement chromatography as first separating step in online two-dimensional liquid chromatography coupled to mass spectrometry analysis of a complex protein sample--the proteome of neutrophils. J CHROMATOGR A, 1232, 288-294. https://doi.org/10.1016/j.chroma.2012.02.029

Vancouver

Trusch M, Tillack K, Kwiatkowski M, Bertsch A, Ahrends R, Kohlbacher O et al. Displacement chromatography as first separating step in online two-dimensional liquid chromatography coupled to mass spectrometry analysis of a complex protein sample--the proteome of neutrophils. J CHROMATOGR A. 2012 Apr 6;1232:288-294. https://doi.org/10.1016/j.chroma.2012.02.029

Bibtex

@article{d1345c2b89fe485ea5dda96a18b639a4,
title = "Displacement chromatography as first separating step in online two-dimensional liquid chromatography coupled to mass spectrometry analysis of a complex protein sample--the proteome of neutrophils",
abstract = "Displacement chromatography provides some advantages over elution chromatography such as the opportunity to enrich trace amounts of molecules and to elute molecules in highest concentrations achievable with liquid chromatography. In a previous study we demonstrated that displacement chromatography is a well-suited alternative to gradient elution in an offline two-dimensional (2D-)LC-MS approach for the analysis of proteomes. In this study we present a method for applying displacement chromatography in an online 2D-LC-MS system including a cation exchange (CEX) column and a reversed phase column. We circumvented the problem of determining the sample capacity of the CEX column by repeated injection (pulses) of sample aliquots monitored by an LC-MS analysis of each flow-through fraction of the CEX column. Elution of tryptic peptides from the CEX column was achieved by repeated injection (pulses) of the displacer spermine. Pulsed displacer injections offer the advantage through physical separation of preventing post-column mixing of already separated compounds. As a proof of principle we analyzed the cytosolic proteome of human neutrophils.",
keywords = "Humans, Peptide Fragments/analysis, Proteomics/*methods, Proteome/*analysis, Chromatography, Ion Exchange/instrumentation/*methods, Chromatography, Reverse-Phase/instrumentation/*methods, Cytosol/chemistry, Databases, Protein, Mass Spectrometry/methods, Neutrophils/chemistry, Humans, Peptide Fragments/analysis, Proteomics/*methods, Proteome/*analysis, Chromatography, Ion Exchange/instrumentation/*methods, Chromatography, Reverse-Phase/instrumentation/*methods, Cytosol/chemistry, Databases, Protein, Mass Spectrometry/methods, Neutrophils/chemistry",
author = "Maria Trusch and Kati Tillack and Marcel Kwiatkowski and Andreas Bertsch and Robert Ahrends and Oliver Kohlbacher and Roland Martin and Mireia Sospedra and Hartmut Schl{\"u}ter",
note = "Copyright {\textcopyright} 2012 Elsevier B.V. All rights reserved.",
year = "2012",
month = apr,
day = "6",
doi = "10.1016/j.chroma.2012.02.029",
language = "English",
volume = "1232",
pages = "288--294",
journal = "J CHROMATOGR A",
issn = "0021-9673",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Displacement chromatography as first separating step in online two-dimensional liquid chromatography coupled to mass spectrometry analysis of a complex protein sample--the proteome of neutrophils

AU - Trusch, Maria

AU - Tillack, Kati

AU - Kwiatkowski, Marcel

AU - Bertsch, Andreas

AU - Ahrends, Robert

AU - Kohlbacher, Oliver

AU - Martin, Roland

AU - Sospedra, Mireia

AU - Schlüter, Hartmut

N1 - Copyright © 2012 Elsevier B.V. All rights reserved.

PY - 2012/4/6

Y1 - 2012/4/6

N2 - Displacement chromatography provides some advantages over elution chromatography such as the opportunity to enrich trace amounts of molecules and to elute molecules in highest concentrations achievable with liquid chromatography. In a previous study we demonstrated that displacement chromatography is a well-suited alternative to gradient elution in an offline two-dimensional (2D-)LC-MS approach for the analysis of proteomes. In this study we present a method for applying displacement chromatography in an online 2D-LC-MS system including a cation exchange (CEX) column and a reversed phase column. We circumvented the problem of determining the sample capacity of the CEX column by repeated injection (pulses) of sample aliquots monitored by an LC-MS analysis of each flow-through fraction of the CEX column. Elution of tryptic peptides from the CEX column was achieved by repeated injection (pulses) of the displacer spermine. Pulsed displacer injections offer the advantage through physical separation of preventing post-column mixing of already separated compounds. As a proof of principle we analyzed the cytosolic proteome of human neutrophils.

AB - Displacement chromatography provides some advantages over elution chromatography such as the opportunity to enrich trace amounts of molecules and to elute molecules in highest concentrations achievable with liquid chromatography. In a previous study we demonstrated that displacement chromatography is a well-suited alternative to gradient elution in an offline two-dimensional (2D-)LC-MS approach for the analysis of proteomes. In this study we present a method for applying displacement chromatography in an online 2D-LC-MS system including a cation exchange (CEX) column and a reversed phase column. We circumvented the problem of determining the sample capacity of the CEX column by repeated injection (pulses) of sample aliquots monitored by an LC-MS analysis of each flow-through fraction of the CEX column. Elution of tryptic peptides from the CEX column was achieved by repeated injection (pulses) of the displacer spermine. Pulsed displacer injections offer the advantage through physical separation of preventing post-column mixing of already separated compounds. As a proof of principle we analyzed the cytosolic proteome of human neutrophils.

KW - Humans

KW - Peptide Fragments/analysis

KW - Proteomics/methods

KW - Proteome/analysis

KW - Chromatography, Ion Exchange/instrumentation/methods

KW - Chromatography, Reverse-Phase/instrumentation/methods

KW - Cytosol/chemistry

KW - Databases, Protein

KW - Mass Spectrometry/methods

KW - Neutrophils/chemistry

KW - Humans

KW - Peptide Fragments/analysis

KW - Proteomics/methods

KW - Proteome/analysis

KW - Chromatography, Ion Exchange/instrumentation/methods

KW - Chromatography, Reverse-Phase/instrumentation/methods

KW - Cytosol/chemistry

KW - Databases, Protein

KW - Mass Spectrometry/methods

KW - Neutrophils/chemistry

U2 - 10.1016/j.chroma.2012.02.029

DO - 10.1016/j.chroma.2012.02.029

M3 - SCORING: Journal article

C2 - 22391494

VL - 1232

SP - 288

EP - 294

JO - J CHROMATOGR A

JF - J CHROMATOGR A

SN - 0021-9673

ER -