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Titolo:
Packed capillary reversed-phase liquid chromatography with high-performance electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry for proteomics
Autore:
Shen, YF; Zhao, R; Belov, ME; Conrads, TP; Anderson, GA; Tang, KQ; Pasa-Tolic, L; Veenstra, TD; Lipton, MS; Udseth, HR; Smith, RD;
Indirizzi:
Battelle Mem Inst, Pacific NW Natl Labs, Environm Mol Sci Lab, Richland, WA 99352 USA Battelle Mem Inst Richland WA USA 99352 l Sci Lab, Richland, WA 99352 USA
Titolo Testata:
ANALYTICAL CHEMISTRY
fascicolo: 8, volume: 73, anno: 2001,
pagine: 1766 - 1775
SICI:
0003-2700(20010415)73:8<1766:PCRLCW>2.0.ZU;2-I
Fonte:
ISI
Lingua:
ENG
Soggetto:
2-DIMENSIONAL GEL-ELECTROPHORESIS; PROTEIN IDENTIFICATION; MIXTURES; COLUMNS; ISOTACHOPHORESIS; TECHNOLOGIES; PEPTIDES; UTILITY; TAGS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Physical, Chemical & Earth Sciences
Citazioni:
44
Recensione:
Indirizzi per estratti:
Indirizzo: Smith, RD Battelle Mem Inst, Pacific NW Natl Labs, Environm Mol Sci Lab, Richland, WA 99352 USA Battelle Mem Inst Richland WA USA 99352 Richland, WA 99352 USA
Citazione:
Y.F. Shen et al., "Packed capillary reversed-phase liquid chromatography with high-performance electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry for proteomics", ANALYT CHEM, 73(8), 2001, pp. 1766-1775

Abstract

In this study, high-efficiency packed capillary reversed-phase liquid chromatography (RPLC) coupled on-line with high-performance Fourier transform ion cyclotron resonance (FTICR) mass spectrometry has been investigated for the characterization of complex cellular proteolytic digests. Long capillary columns (80-cm) packed with small (3-mum) C18 bonded particles provided atotal peak capacity of similar to 1000 for cellular proteolytic polypeptides when interfaced with an ESI-FTICR mass spectrometer under composition gradient conditions at a pressure of 10 000 psi, Large quantities of cellularproteolytic digests (e.g,, 500 mug) could be loaded onto packed capillaries of 150-mum inner diameter without a significant loss of separation efficiency. Precolumns with suitable inner diameters were found useful for improving the elution reproducibility without a significant loss of separation quality. Porous particle packed capillaries were found to provide better results than those containing nonporous particles because of their higher sample capacity. Two-dimensional analyses from the combination of packed capillary RPLC with high-resolution FTICR yield a combined capacity for separations of >1 million polypeptide components and simultaneously provided information for the identification of the separated components based upon the accurate mass tag concept previously described.

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Documento generato il 24/09/20 alle ore 04:27:36