Capillary Electrophoresis as a Second Dimension to Isoelectric Focusing for Peptide Separation

• Published in: Analytical chemistry (Washington) 2007-08, Vol.79 (15), p.5949-5955
• Main Authors: Busnel, Jean-Marc, Lion, Niels, Girault, Hubert H
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Animals; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography; Chromatography, Liquid - methods; Cytochromes c - isolation & purification; Cytochromes c - metabolism; Electrophoresis, Capillary - methods; Exact sciences and technology; Isoelectric Focusing - methods; Lactoglobulins - isolation & purification; Lactoglobulins - metabolism; Myoglobin - isolation & purification; Myoglobin - metabolism; Other chromatographic methods; Peptide Fragments - isolation & purification; Peptide Fragments - metabolism; Peptides; Proteins; Serum Albumin, Bovine - isolation & purification; Serum Albumin, Bovine - metabolism; Trypsin - metabolism
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac070493r

Capillary zone electrophoresis and carrier ampholytes based capillary electrophoresis have been used as a second separation step to Off-Gel isoelectric focusing for the analysis of complex peptide mixtures. A tryptic digest of four proteins (bovine serum albumin, β-lactoglobulin, horse myoglobin, cytochrome c) has been chosen as a peptide test mixture. After assessment of different modes of capillary electrophoresis as a second dimension to Off-Gel isoelectric focusing, the optimized two-dimensional platforms provide a degree of orthogonality comparable to state-of-the-art multidimensional liquid chromatography systems as well as a practical peak capacity above 700.