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Titolo:
Comparison of the capabilities of liquid isoelectric focusing-one-dimensional nonporous silica reversed-phase liquid chromatography-electrospray ionization time-of-flight mass spectrometry and liquid isoelectric focusing-one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis massmapping for the analysis of intact protein molecular masses
Autore:
Wall, DB; Parus, SJ; Lubman, DM;
Indirizzi:
Univ Michigan, Dept Chem, Ann Arbor, MI 48109 USA Univ Michigan Ann ArborMI USA 48109 , Dept Chem, Ann Arbor, MI 48109 USA
Titolo Testata:
JOURNAL OF CHROMATOGRAPHY B
fascicolo: 1-2, volume: 763, anno: 2001,
pagine: 139 - 148
SICI:
1387-2273(20011105)763:1-2<139:COTCOL>2.0.ZU;2-V
Fonte:
ISI
Lingua:
ENG
Soggetto:
ASSISTED-LASER-DESORPTION/IONIZATION; HUMAN ERYTHROLEUKEMIA-CELLS; IDENTIFICATION; DIGESTION;
Keywords:
mass mapping; molecular mass; proteins;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
29
Recensione:
Indirizzi per estratti:
Indirizzo: Lubman, DM Univ Michigan, Dept Chem, 930 N Univ Ave, Ann Arbor, MI 48109 USA Univ Michigan 930 N Univ Ave Ann Arbor MI USA 48109 I 48109 USA
Citazione:
D.B. Wall et al., "Comparison of the capabilities of liquid isoelectric focusing-one-dimensional nonporous silica reversed-phase liquid chromatography-electrospray ionization time-of-flight mass spectrometry and liquid isoelectric focusing-one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis massmapping for the analysis of intact protein molecular masses", J CHROMAT B, 763(1-2), 2001, pp. 139-148

Abstract

Nonporous silica reversed-phase HPLC coupled to electrospray ionization with on-line time-of-flight mass spectrometric detection (NPS-RP-HPLC-ESI-TOF-MS) is shown to be an effective liquid phase method for obtaining the molecular masses of proteins from pH fractionated cellular lysates where the method is capable of generating the same banding patterns typically observed using gel phase one-dimensional sodium dodecyl sulfatepolyacrylamide gel electrophoresis. The liquid-phase mass spectrometry-based method provides a mass accuracy of at least 150 ppm, with 4000 mass resolution and provides improved sensitivity as the protein molecular mass (MW) decreases. The liquidand gel phase methods are shown to be complementary in terms of their massrange but the liquid phase method has the advantage over the gel method inthat the analysis times are 50 times shorter, the mass accuracy is 70 times better and the resolution is 130 times higher. The liquid phase method isshown to be more effective for detection of proteins below 40 kDa, while the gel phase separation can access many more proteins, including more hydrophobic proteins, at increasing MW. (C) 2001 Elsevier Science B.V. All rights reserved.

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Documento generato il 05/12/20 alle ore 01:12:01