Comprehensive two‐dimensional chromatography and capillary electrophoresis coupled with tandem time‐of‐flight mass spectrometry for high‐speed proteome analysis
- 14 July 2004
- journal article
- research article
- Published by Wiley in Electrophoresis
- Vol. 25 (14) , 2374-2383
- https://doi.org/10.1002/elps.200405956
Abstract
A comprehensive two‐dimensional capillary liquid chromatography and capillary zone electrophoresis system coupled with tandem matrix assisted laser desorption/ionization‐time of flight‐time of flight‐mass spectrometry (MALDI‐TOF‐TOF‐MS) proteomics analyzer is presented. Protein/peptide samples were separated by capillary high‐performance liquid chromatography (cHPLC). The effluents from cHPLC (the first dimension) were continuously transferred into capillary zone electrophoresis (CZE, the second dimension) through a novel valve‐free hydrodynamic sampling interface. The CZE effluents were mixed with α‐cyano‐4‐hydroxycinnamic acid (CHCA) matrix sheath flow via CE‐MALDI interface, and then directly deposited on the MALDI target at a 3 s time‐interval for further MS analysis. The high efficiency of the overall system was demonstrated by analysis of proteins in D20 (human hepatocellular carcinoma model in nude mice with high metastatic potential) liver cancer tissue. More than 300 proteins were identified, which proved the system potential for high‐throughput analysis and application in proteomics.Keywords
This publication has 22 references indexed in Scilit:
- Comprehensive two-dimensional separation system by coupling capillary reverse-phase liquid chromatography to capillary isoelectric focusing for peptide and protein mapping with laser-induced fluorescence detectionElectrophoresis, 2003
- Comprehensive two‐dimensional separations based on capillary high‐performance liquid chromatography and microchip electrophoresisElectrophoresis, 2003
- On-Line Concentration of Proteins and Peptides in Capillary Zone Electrophoresis with an Etched Porous JointAnalytical Chemistry, 2002
- A CE−MALDI Interface Based on the Use of Prestructured Sample SupportsAnalytical Chemistry, 2001
- A Transparent Flow Gating Interface for the Coupling of Microcolumn LC with CZE in a Comprehensive Two-Dimensional SystemAnalytical Chemistry, 1997
- Off-Line Coupling of Capillary Electrophoresis and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass SpectrometryAnalytical Chemistry, 1995
- Rapid comprehensive two-dimensional separations of peptides via RPLC-optically gated capillary zone electrophoresisAnalytical Chemistry, 1995
- Separation of Nanoliter Samples of Biological Amines by a Comprehensive Two-Dimensional Microcolumn Liquid Chromatography SystemAnalytical Chemistry, 1995
- Study of zone broadening in optically gated high-speed capillary electrophoresisAnalytical Chemistry, 1993
- Cellulose acetate-coated porous polymer joint for capillary zone electrophoresisAnalytical Chemistry, 1992