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Rapid selection of peptide containing fractions in off‐line 2‐D HPLC in shotgun proteome analysis by screening with MALDI TOF MS |
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| Authors: | Maria Lasaosa Thomas Jakoby Nathanaël Delmotte Christian G. Huber Elmar Heinzle Andreas Tholey |
| Affiliation: | 1. Technische Biochemie, Universit?t des Saarlandes, Saarbrücken, Germany;2. Center for Bioinformatics – Junior Research Group for Protein–Protein Interactions and Computational Proteomics, Universit?t des Saarlandes, Saarbrücken, Germany;3. Division for Systematic Proteome Research, Institute for Experimental Medicine, Christian‐Albrechts‐University, Kiel, Germany;4. Instrumental Analysis and Bioanalysis, Universit?t des Saarlandes, Saarbrücken, Germany;5. Department of Molecular Biology, Division of Chemistry and Bioanalytics, University of Salzburg, Salzburg, Austria |
| Abstract: | A simple and fast screening method for the selection of fractions of first dimension separation to be analyzed in second dimension‐MS/MS experiments in offline multidimensional liquid chromatographic separation schemes for shotgun proteome analysis was developed. The method is based on the measurement of total peptide content of the first dimension fractions by MALDI MS and was established using a tryptic digest of a bacterial proteome. The results of the screening process were in good agreement with those obtained in a detailed proteome analysis performed by RP×ion‐pair RP‐MALDI TOF/TOF MS analysis. The method supports a straightforward planning of experiments, also enabling a reduction of overall measurement time in shotgun proteome analysis. |
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