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.

Basic information

Original name

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

Authors

REJTAR, Tomas (203 Czech Republic), Hu PING (156 China), Peter JUHASZ (840 United States of America), Jennifer M. CAMPBELL (840 United States of America), Marvin L. VESTAL (840 United States of America), Jan PREISLER (203 Czech Republic, guarantor) and Barry, L. KARGER (840 United States of America)

Edition

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

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10406 Analytical chemistry

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14310/02:00008546

Organization unit

Faculty of Science

UT WoS

000178238600008

Keywords in English

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

Abstract

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 and 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, vol. 1, No 2, p. 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 and 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, vol.~1, No~2, p.~171-179. ISSN~1535-3893.

Detailed Information on Publication Record