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Rapid identification of differentiation markers from whole epithelial cells by matrix‐assisted laser desorption/ionisation time‐of‐flight mass spectrometry and statistical analysis
Author(s) -
MarvinGuy Laure F.,
Duncan Peter,
Wagnière Sandrine,
Antille Nicolas,
Porta Nadine,
Affolter Michael,
Kussmann Martin
Publication year - 2008
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.3479
Subject(s) - chemistry , mass spectrometry , cellular differentiation , chromatography , ion suppression in liquid chromatography–mass spectrometry , matrix assisted laser desorption/ionization , tandem mass spectrometry , matrix (chemical analysis) , desorption , biochemistry , organic chemistry , adsorption , gene
Abstract Matrix‐assisted laser desorption/ionisation time‐of‐flight mass spectrometry (MALDI‐TOFMS) was applied to identify markers for cellular differentiation. The differentiation of a human colon epithelial carcinoma T84 cell line was monitored over a period of 28 days by transepithelial electrical resistance (TER) measurements, alkaline phosphatase (AP) assay, and MALDI‐TOF mass spectral fingerprints combined with statistical analysis. MALDI‐MS generated specific mass spectral fingerprints characteristic of cell differentiation. Twenty‐two ions were selected as diagnostic signals of fully differentiated T84 cells. Ten protein ion signals, detected by MALDI‐MS and validated by statistical analysis, were proposed as T84 cell differentiation markers. Among these signals, ubiquitin was identified as a T84 cell differentiation marker by nanospray liquid chromatography/tandem mass spectrometry (nanoLC/MS/MS). Moreover, depending on the concentration of the cells seeded on the growth support, it was possible to predict the timing of the exponential phase and of cellular differentiation by MALDI‐MS‐derived marker ions. MALDI‐TOFMS was compared to other methods for the determination of cellular differentiation: TER measurements are rapid but yield limited information as to the cellular differentiation state. AP assays are more specific for the differentiation state but take more time. By contrast, MALDI‐MS has been found to be a fast, sensitive and precise method for cell differentiation assessment and provides the opportunity for multiplexing and high throughput. Moreover, the consumable costs per assay are very low. Copyright © 2008 John Wiley & Sons, Ltd.