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Title: Identification of mammalian cell lines using MALDI-TOF and LC-ESI-MS/MS mass spectrometry

Abstract

Direct mass spectrometric analysis of complex biological samples is becoming an increasingly useful technique in the field of proteomics. Matrix-assisted laser desorption/ionization mass spectroscopy (MALDI-MS) is a rapid and sensitive analytical tool well suited for obtaining molecular weights of peptides and proteins from complex samples. Here, a fast and simple approach to cellular protein profiling is described in which mammalian cells are lysed directly in the MALDI matrix 2,5-dihydroxybenzoic acid (DHB) and mass analyzed using MALDI-time of flight (TOF). Using the unique MALDI mass spectral “fingerprint” generated in these analyses, it is possible to differentiate among several different mammalian cell lines. A number of techniques, including MALDI-post source decay (PSD), MALDI tandem time-of-flight (TOF-TOF), MALDI-Fourier transform ion cyclotron resonance (FTICR), and nanoflow liquid chromatography followed by electrospray ionization and tandem mass spectrometry (LC-ESI-MS/MS) were employed to attempt to identify the proteins represented in the MALDI spectra. Performing a tryptic digestion of the supernatant of the cells lysed in DHB with subsequent LC-ESI-MS/MS analysis was by far the most successful method to identify proteins.

Authors:
; ; ;
Publication Date:
OSTI Identifier:
22774061
Resource Type:
Journal Article
Journal Name:
Journal of the American Society for Mass Spectrometry
Additional Journal Information:
Journal Volume: 17; Journal Issue: 4; Other Information: Copyright (c) 2006 American Society for Mass Spectrometry; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 1044-0305
Country of Publication:
United States
Language:
English
Subject:
46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND TECHNOLOGY; AUGMENTATION; DESORPTION; FOURIER TRANSFORMATION; ION CYCLOTRON-RESONANCE; IONIZATION; LASER RADIATION; LIQUID COLUMN CHROMATOGRAPHY; MASS; MASS SPECTROSCOPY; MOLECULAR WEIGHT; SPECTRA; TIME-OF-FLIGHT METHOD

Citation Formats

Zhang, Xu, Scalf, Mark, Berggren, Travis W., Westphall, Michael S., and Smith, Lloyd M., E-mail: smith@chem.wisc.edu. Identification of mammalian cell lines using MALDI-TOF and LC-ESI-MS/MS mass spectrometry. United States: N. p., 2006. Web. doi:10.1016/J.JASMS.2005.12.007.
Zhang, Xu, Scalf, Mark, Berggren, Travis W., Westphall, Michael S., & Smith, Lloyd M., E-mail: smith@chem.wisc.edu. Identification of mammalian cell lines using MALDI-TOF and LC-ESI-MS/MS mass spectrometry. United States. doi:10.1016/J.JASMS.2005.12.007.
Zhang, Xu, Scalf, Mark, Berggren, Travis W., Westphall, Michael S., and Smith, Lloyd M., E-mail: smith@chem.wisc.edu. Sat . "Identification of mammalian cell lines using MALDI-TOF and LC-ESI-MS/MS mass spectrometry". United States. doi:10.1016/J.JASMS.2005.12.007.
@article{osti_22774061,
title = {Identification of mammalian cell lines using MALDI-TOF and LC-ESI-MS/MS mass spectrometry},
author = {Zhang, Xu and Scalf, Mark and Berggren, Travis W. and Westphall, Michael S. and Smith, Lloyd M., E-mail: smith@chem.wisc.edu},
abstractNote = {Direct mass spectrometric analysis of complex biological samples is becoming an increasingly useful technique in the field of proteomics. Matrix-assisted laser desorption/ionization mass spectroscopy (MALDI-MS) is a rapid and sensitive analytical tool well suited for obtaining molecular weights of peptides and proteins from complex samples. Here, a fast and simple approach to cellular protein profiling is described in which mammalian cells are lysed directly in the MALDI matrix 2,5-dihydroxybenzoic acid (DHB) and mass analyzed using MALDI-time of flight (TOF). Using the unique MALDI mass spectral “fingerprint” generated in these analyses, it is possible to differentiate among several different mammalian cell lines. A number of techniques, including MALDI-post source decay (PSD), MALDI tandem time-of-flight (TOF-TOF), MALDI-Fourier transform ion cyclotron resonance (FTICR), and nanoflow liquid chromatography followed by electrospray ionization and tandem mass spectrometry (LC-ESI-MS/MS) were employed to attempt to identify the proteins represented in the MALDI spectra. Performing a tryptic digestion of the supernatant of the cells lysed in DHB with subsequent LC-ESI-MS/MS analysis was by far the most successful method to identify proteins.},
doi = {10.1016/J.JASMS.2005.12.007},
journal = {Journal of the American Society for Mass Spectrometry},
issn = {1044-0305},
number = 4,
volume = 17,
place = {United States},
year = {2006},
month = {4}
}