Capillary electrophoresis-mass spectrometry for the analysis of intact proteins.
Developments in the fields of protein chemistry, proteomics and biotechnology have increased the demand for suitable analytical techniques for the analysis of intact proteins. In 1989, capillary electrophoresis (CE) was combined with mass spectrometry (MS) for the first time and its potential usefulness for the analysis of intact (i.e. non-digested) proteins was shown. This article provides an overview of the applications of CE-MS within the field of intact protein analysis. The principles of the applied CE modes and ionization techniques used for CE-MS of intact proteins are shortly described. It is shown that separations are predominantly carried out by capillary zone electrophoresis and capillary isoelectric focusing, whereas electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI) are the most popular ionization techniques used for interfacing. The combination of CE with inductively coupled plasma (ICP) MS for the analysis of metalloproteins is also discussed. The various CE-MS combinations are systematically outlined and tables provide extensive overviews of the applications of each technique for intact protein analysis. Selected examples are given to illustrate the usefulness of the CE-MS techniques. Examples include protein isoform assignment, single cell analysis, metalloprotein characterization, proteomics and biomarker screening. Finally, chip-based electrophoresis combined with MS is shortly treated and some of its applications are described. It is concluded that CE-MS represents a powerful tool for the analysis of intact proteins yielding unique separations and information.
Haselberg
Rob
R
Department of Biomedical Analysis, Utrecht University, 3508 TB Utrecht, The Netherlands. r.haselberg@uu.nl
de Jong
Gerhardus J
GJ
Somsen
Govert W
GW
D015415:Biomarkers / Q000032:analysis; D002021:Buffers; D053801:Capillary Electrochromatography / Q000379:methods; D020099:Coated Materials, Biocompatible; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D007525:Isoelectric Focusing / Q000295:instrumentation / Q000379:methods; D008667:Metalloproteins / Q000032:analysis; D017510:Protein Folding; D020033:Protein Isoforms / Q000032:analysis; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013054:Spectrophotometry, Atomic / Q000379:methods
D015415:Biomarkers; D002021:Buffers; D020099:Coated Materials, Biocompatible; D008667:Metalloproteins; D020033:Protein Isoforms; D011506:Proteins