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2013 | 11 | 1 | 25-34
Tytuł artykułu

Fully automated chip-based nanoelectrospray combined with electron transfer dissociation for high throughput top-down proteomics

Treść / Zawartość
Warianty tytułu
Języki publikacji
EN
Abstrakty
EN
The conventional protocol for protein identification by electrospray ionization mass spectrometry (MS) is based on enzymatic digestion which renders peptides to be analyzed by liquid chromatography-MS and collision-induced dissociation (CID) multistage MS, in the so-called bottom-up approach. Though this method has brought a significant progress to the field, many limitations, among which, the low throughput and impossibility to characterize in detail posttranslational modifications in terms of site(s) and structure, were reported. Therefore, the research is presently focused on the development of procedures for efficient top-down fragmentation of intact protein ions. In this context, we developed here an approach combining fully automated chip-based-nanoelectrospray ionisation (nanoESI), performed on a NanoMate robot, with electron transfer dissociation (ETD) for peptide and top-down protein sequencing and identification. This advanced analytical platform, integrating robotics, microfluidics technology, ETD and alternate ETD/CID, was tested and found ideally suitable for structural investigation of peptides and modified/functionalized peptides as well as for top-down analysis of medium size proteins by tandem MS experiments of significantly increased throughput and sensitivity. The obtained results indicate that NanoMate-ETD and ETD/CID may represent a viable alternative to the current MS strategies, with potential to develop into a method of routine use for high throughput top-down proteomics.
Wydawca

Czasopismo
Rocznik
Tom
11
Numer
1
Strony
25-34
Opis fizyczny
Daty
wydano
2013-01-01
online
2012-10-24
Twórcy
  • “Aurel Vlaicu” University of Arad
  • “Aurel Vlaicu” University of Arad
  • “Aurel Vlaicu” University of Arad
  • University of Konstanz
autor
  • Chemistry Institute of Romanian Academy
autor
Bibliografia
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Typ dokumentu
Bibliografia
Identyfikatory
Identyfikator YADDA
bwmeta1.element.-psjd-doi-10_2478_s11532-012-0130-2
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