Abstract
The molecular scanner combines protein separation using gel electrophoresis with peptide mass fingerprinting (PMF) techniques to identify proteins in a highly automated manner. Proteins separated in a 2-dimensional polyacrylamide gel (2D-PAGE) are digested ‘in parallel’ and transferred onto a membrane keeping their relative positions. The membrane is then sprayed with a matrix and inserted into a matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometer, which measures a peptide mass fingerprint at each site on the scanned grid. First, visualization of PMF data allows surveying all fingerprints at once and provides very useful information on the presence of chemical noise. Chemical noise is shown to be a potential source for erroneous identifications and is therefore purged from the mass fingerprints. Then, the correlation between neighboring spectra is used to recalibrate the peptide masses. Finally, a method that clusters peptide masses according to the similarity of the spatial distributions of their signal intensities is presented. This method allows discarding many of the false positives that usually go along with PMF identifications and allows identifying many weakly expressed proteins present in the gel.
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Muller, M., Gras, R., Appel, R.D., Bienvenut, W.V., Hochstrasser, D.F. (2005). Signal Traitment and Virtual Images Production (2/2). In: Bienvenut, W.V. (eds) Acceleration and Improvement of Protein Identification by Mass Spectrometry. Springer, Dordrecht. https://doi.org/10.1007/1-4020-3319-2_5
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DOI: https://doi.org/10.1007/1-4020-3319-2_5
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