Off-Line Coupling of High-Resolution Capillary Electrophoresis
to MALDI-TOF and TOF/TOF MS

J 2002

Off-Line Coupling of High-Resolution Capillary Electrophoresis to MALDI-TOF and TOF/TOF MS

REJTAR, Tomas; Hu PING; Peter JUHASZ; Jennifer M. CAMPBELL; Marvin L. VESTAL et. al.

Základní údaje

Originální název

Off-Line Coupling of High-Resolution Capillary Electrophoresis to MALDI-TOF and TOF/TOF MS

Autoři

REJTAR, Tomas (203 Česká republika); Hu PING (156 Čína); Peter JUHASZ (840 Spojené státy); Jennifer M. CAMPBELL (840 Spojené státy); Marvin L. VESTAL (840 Spojené státy); Jan PREISLER (203 Česká republika, garant) a Barry, L. KARGER (840 Spojené státy)

Vydání

Journal of Proteome Research, Washington, USA, ACS, 2002, 1535-3893

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Kód RIV

RIV/00216224:14310/02:00008546

Organizační jednotka

Přírodovědecká fakulta

UT WoS

000178238600008

Klíčová slova anglicky

MALDI; mass spectrometry; capillary electrophoresis; proteomics; proteins; peptides; interface;

Štítky

Capillary electrophoresis, interface, MALDI, mass spectrometry, peptides, proteins, proteomics

Změněno: 27. 6. 2009 23:58, prof. Mgr. Jan Preisler, Ph.D.

Anotace

V originále

High-resolution capillary electrophoresis has been coupled to MALDI-TOF and TOF/TOF MS through off-line vacuum deposition onto standard stainless steel MALDI targets. This off-line approach allowed the decoupling of the separation from the MS analysis, thus allowing each to be independently optimized in terms of time. Using BSA tryptic digest as a model sample, the deposited streaks, roughly 100-ím wide, were first analyzed in the MS mode, consuming only a fraction of the sample. After data analysis, segments of the deposited trace, containing unidentified peptides, as well as several species chosen for sequence confirmation, were reanalyzed in the MS/MS mode using MALDI-TOF/TOF MS. Additionally, it is shown that the shot-to-shot reproducibility of the vacuum-deposited trace (5% RSD) is 1 order of magnitude lower than that found for the standard dried droplet method. Moreover, a linear dependence of signal intensities (relative to an internal standard) over 3 orders of magnitude was found for a peptide sample with concentrations ranging from 1 to 1000 nM. This paper demonstrates the potential of off-line coupling of high-resolution separations to MALDI-MS and MALDI-MS/MS using vacuum deposition for the analysis of complex peptide mixtures from protein digests.

Citovat

REJTAR, Tomas; Hu PING; Peter JUHASZ; Jennifer M. CAMPBELL; Marvin L. VESTAL; Jan PREISLER a Barry, L. KARGER. Off-Line Coupling of High-Resolution Capillary Electrophoresis to MALDI-TOF and TOF/TOF MS. Journal of Proteome Research. Washington, USA: ACS, 2002, roč. 1, č. 2, s. 171-179. ISSN 1535-3893.

@article{397051,
   author = {Rejtar, Tomas and Ping, Hu and Juhasz, Peter and Campbell, Jennifer M. and Vestal, Marvin L. and Preisler, Jan and Karger, Barry, L.},
   article_location = {Washington, USA},
   article_number = {2},
   keywords = {MALDI; mass spectrometry; capillary electrophoresis; proteomics; proteins; peptides; interface;},
   language = {eng},
   issn = {1535-3893},
   journal = {Journal of Proteome Research},
   title = {Off-Line Coupling of High-Resolution Capillary Electrophoresis to MALDI-TOF and TOF/TOF MS},
   volume = {1},
   year = {2002}
}

TY  - JOUR
ID  - 397051
AU  - Rejtar, Tomas - Ping, Hu - Juhasz, Peter - Campbell, Jennifer M. - Vestal, Marvin L. - Preisler, Jan - Karger, Barry, L.
PY  - 2002
TI  - Off-Line Coupling of High-Resolution Capillary Electrophoresis to MALDI-TOF and TOF/TOF MS
JF  - Journal of Proteome Research
VL  - 1
IS  - 2
SP  - 171
EP  - 171
PB  - ACS
SN  - 15353893
KW  - MALDI
KW  - mass spectrometry
KW  - capillary electrophoresis
KW  - proteomics
KW  - proteins
KW  - peptides
KW  - interface;
N2  - High-resolution capillary electrophoresis has been coupled to MALDI-TOF and TOF/TOF MS through off-line vacuum deposition onto standard stainless steel MALDI targets. This off-line approach allowed the decoupling of the separation from the MS analysis, thus allowing each to be independently optimized in terms of time. Using BSA tryptic digest as a model sample, the deposited streaks, roughly 100-ím wide, were first analyzed in the MS mode, consuming only a fraction of the sample. After data analysis, segments of the deposited trace, containing unidentified peptides, as well as several species chosen for sequence confirmation, were reanalyzed in the MS/MS mode using MALDI-TOF/TOF MS. Additionally, it is shown that the shot-to-shot reproducibility of the vacuum-deposited trace (5% RSD) is 1 order of magnitude lower than that found for the standard dried droplet method. Moreover, a linear dependence of signal intensities (relative to an internal standard) over 3 orders of magnitude was found for a peptide sample with concentrations ranging from 1 to 1000 nM. This paper demonstrates the potential of off-line coupling of high-resolution separations to MALDI-MS and MALDI-MS/MS using vacuum deposition for the analysis of complex peptide mixtures from protein digests.
ER  -

REJTAR, Tomas; Hu PING; Peter JUHASZ; Jennifer M. CAMPBELL; Marvin L. VESTAL; Jan PREISLER a Barry, L. KARGER. Off-Line Coupling of High-Resolution Capillary Electrophoresis to MALDI-TOF and TOF/TOF MS. \textit{Journal of Proteome Research}. Washington, USA: ACS, 2002, roč.~1, č.~2, s.~171-179. ISSN~1535-3893.

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