MALDI imaging and profiling MS of higher mass proteins from tissue |
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Authors: | Alexandra van Remoortere René J. M. van Zeijl Nico van den Oever Julien Franck Rémi Longuespée Maxence Wisztorski Michel Salzet André M. Deelder Isabelle Fournier Liam A. McDonnell |
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Affiliation: | 1.Biomolecular Mass Spectrometry Unit, Department of Parasitology,Leiden University Medical Center,Leiden,The Netherlands;2.Hogeschool Leiden,Leiden,The Netherlands;3.Université de Lillel, CNRS-FRE 3249, MALDI Imaging Team,Laboratoire de Neuroimmunologie et Neurochimie Evolutives,Villeneuve d’Ascq,France |
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Abstract: | MALDI imaging and profiling mass spectrometry of proteins typically leads to the detection of a large number of peptides and small proteins but is much less successful for larger proteins: most ion signals correspond to proteins of m/z < 25,000. This is a severe limitation as many proteins, including cytokines, growth factors, enzymes, and receptors have molecular weights exceeding 25 kDa. The detector technology typically used for protein imaging, a microchannel plate, is not well suited to the detection of high m/z ions and is prone to detector saturation when analyzing complex mixtures. Here we report increased sensitivity for higher mass proteins by using the CovalX high mass HM1 detector (Zurich, Switzerland), which has been specifically designed for the detection of high mass ions and which is much less prone to detector saturation. The results demonstrate that a range of different sample preparation strategies enable higher mass proteins to be analyzed if the detector technology maintains high detection efficiency throughout the mass range. The detector enables proteins up to 70 kDa to be imaged, and proteins up to 110 kDa to be detected, directly from tissue, and indicates new directions by which the mass range amenable to MALDI imaging MS and MALDI profiling MS may be extended. |
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