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

Journal Article · Sat Apr 15 04:00:00 EDT 2006 · Journal of the American Society for Mass Spectrometry

DOI:https://doi.org/10.1016/J.JASMS.2005.12.007· OSTI ID:22774061

Zhang, Xu; Scalf, Mark; Berggren, Travis W.; Westphall, Michael S.

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.

• OSTI ID: 22774061
• Journal Information: Journal of the American Society for Mass Spectrometry, Journal Name: Journal of the American Society for Mass Spectrometry Journal Issue: 4 Vol. 17; ISSN 1044-0305; ISSN JAMSEF
• Country of Publication: United States
• Language: English

Related Subjects

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