Identification of proteins by capillary electrophoresis-tandem mass spectrometry. Evaluation of an on-line solid-phase extraction device.
Capillary electrophoresis-tandem mass spectrometry has been used successfully for the analysis of complex peptide mixtures. The method is limited by a relatively high concentration limit of detection and by matrix effects. Here we describe on-line coupling of a solid-phase microextraction device to a capillary electrophoresis-tandem mass spectrometry system. The performance of the integrated instrument was evaluated for the identification of proteins by their amino acid sequence. We report that the concentration limit of detection was improved at least 1000 fold to the low attomole/microliter range and that matrix effects were minimized by extensive sample clean-up during solid-phase extraction. We demonstrate that the implementation of a solid-phase extraction device significantly enhances capillary electrophoresis-tandem mass spectrometry as a method for the identification of low abundance proteins isolated from high-resolution two-dimensional polyacrylamide gels.
Figeys
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D
Department of Molecular Biotechnology, University of Washington, Seattle 98195-7730, USA.
Ducret
A
A
Aebersold
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R
D000595:Amino Acid Sequence; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional; D005656:Fungal Proteins / Q000032:analysis; D013058:Mass Spectrometry / Q000295:instrumentation* / Q000379:methods; D008832:Microchemistry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis / Q000378:metabolism; D010449:Peptide Mapping; D011506:Proteins / Q000032:analysis*; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000032:analysis; D014357:Trypsin / Q000378:metabolism
D005656:Fungal Proteins; D010446:Peptide Fragments; D011506:Proteins; D012710:Serum Albumin, Bovine; D014357:Trypsin