Shotgun Mass Spectrometry Workflow Combining IEF and LC-MALDI-TOF/TOF

We present a high throughput shotgun mass spectrometry workflow using a bidimensional peptide fractionation procedure consisting of isoelectric focusing and RP-HPLC prior to mass spectrometric analysis, with the aim of optimizing peptide separation and protein identification. As part of the workflow we used the 'Isotope-Coded Protein Labeling' (ICPL) method for accurate relative quantitation of protein expression. Such workflow was successfully applied to a comparative proteome analysis of schizophrenia versus healthy control brain tissues and can be an alternative to proteome researches.