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Protein identification platform utilizing micro dispensing technology interfaced to matrix-assisted laser desorption ionization time-of-flight mass spectrometry

Miliotis, Tasso LU ; Kjellström, Sven LU ; Önnerfjord, Patrik LU orcid ; Nilsson, Johan LU ; Laurell, Thomas LU ; Edholm, Lars Erik and Marko-Varga, György LU (2000) In Journal of Chromatography A 886(1-2). p.99-110
Abstract

An integrated protein microcharacterization/identification platform has been developed. The system has been designed to allow a high flexibility in order to tackle challenging analytical problems. The platform comprises a cooled microautosampler, an integrated system for microcolumn HPLC, and a capillary reversed-phase column that is interfaced to matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) system via a low internal volume flow-through microdispenser. The chromatographic separation is continuously transferred onto a MALDI target plate as discrete spots as the dispenser ejects bursts of droplets of the column effluent in a precise array pattern. A refrigerated microfraction collector was... (More)

An integrated protein microcharacterization/identification platform has been developed. The system has been designed to allow a high flexibility in order to tackle challenging analytical problems. The platform comprises a cooled microautosampler, an integrated system for microcolumn HPLC, and a capillary reversed-phase column that is interfaced to matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) system via a low internal volume flow-through microdispenser. The chromatographic separation is continuously transferred onto a MALDI target plate as discrete spots as the dispenser ejects bursts of droplets of the column effluent in a precise array pattern. A refrigerated microfraction collector was coupled to the outlet of the flow-through microdispenser enabling enrichment and re-analysis of interesting fractions. The use of target plates pre-coated with matrix simplified and increased the robustness of the system. By including a separation step prior to the MALDI-TOF-MS analysis and hereby minimizing suppression effects allowed us to obtain higher sequence coverage of proteins compared to conventional MALDI sample preparation methodology. Additionally, synthetic peptides corresponding to autophosphorylated forms of the tryptic fragment 485-496 (ALGADDSYYTAR) of tyrosine kinase ZAP-70 were identified at sensitivities reaching 150 amol. Copyright (C) 2000 Elsevier Science B.V.

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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Instrumentation, Interfaces, LC-MS, Matrix-assisted laser desorption ionization mass spectrometry, Proteins
in
Journal of Chromatography A
volume
886
issue
1-2
pages
12 pages
publisher
Elsevier
external identifiers
  • scopus:0034698458
  • pmid:10950280
ISSN
0021-9673
DOI
10.1016/S0021-9673(00)00430-1
language
English
LU publication?
yes
id
d6d6af7b-d82f-4bb9-9464-19cb82fcfdc4
date added to LUP
2016-10-14 11:36:26
date last changed
2023-09-12 01:50:53
@article{d6d6af7b-d82f-4bb9-9464-19cb82fcfdc4,
  abstract     = {{<p>An integrated protein microcharacterization/identification platform has been developed. The system has been designed to allow a high flexibility in order to tackle challenging analytical problems. The platform comprises a cooled microautosampler, an integrated system for microcolumn HPLC, and a capillary reversed-phase column that is interfaced to matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) system via a low internal volume flow-through microdispenser. The chromatographic separation is continuously transferred onto a MALDI target plate as discrete spots as the dispenser ejects bursts of droplets of the column effluent in a precise array pattern. A refrigerated microfraction collector was coupled to the outlet of the flow-through microdispenser enabling enrichment and re-analysis of interesting fractions. The use of target plates pre-coated with matrix simplified and increased the robustness of the system. By including a separation step prior to the MALDI-TOF-MS analysis and hereby minimizing suppression effects allowed us to obtain higher sequence coverage of proteins compared to conventional MALDI sample preparation methodology. Additionally, synthetic peptides corresponding to autophosphorylated forms of the tryptic fragment 485-496 (ALGADDSYYTAR) of tyrosine kinase ZAP-70 were identified at sensitivities reaching 150 amol. Copyright (C) 2000 Elsevier Science B.V.</p>}},
  author       = {{Miliotis, Tasso and Kjellström, Sven and Önnerfjord, Patrik and Nilsson, Johan and Laurell, Thomas and Edholm, Lars Erik and Marko-Varga, György}},
  issn         = {{0021-9673}},
  keywords     = {{Instrumentation; Interfaces, LC-MS; Matrix-assisted laser desorption ionization mass spectrometry; Proteins}},
  language     = {{eng}},
  month        = {{07}},
  number       = {{1-2}},
  pages        = {{99--110}},
  publisher    = {{Elsevier}},
  series       = {{Journal of Chromatography A}},
  title        = {{Protein identification platform utilizing micro dispensing technology interfaced to matrix-assisted laser desorption ionization time-of-flight mass spectrometry}},
  url          = {{http://dx.doi.org/10.1016/S0021-9673(00)00430-1}},
  doi          = {{10.1016/S0021-9673(00)00430-1}},
  volume       = {{886}},
  year         = {{2000}},
}