Comprehensive two‐dimensional chromatography and capillary electrophoresis coupled with tandem time‐of‐flight mass spectrometry for high‐speed proteome analysis

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 alpha-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.

Proteome Analysis of Microdissected Formalin-fixed and Paraffin-embedded Tissue Specimens

“…Thus, most tissue proteomic studies have employed multidimensional liquid chromatography separations (Li et al 2004;Zhang et al 2004;Cagney et al 2005;DeSouza et al 2005) and are mainly based on analysis of entire tissue sections instead of targeted subpopulations of microdissectionderived cells. However, the heterogeneous nature of most tissues, as well as the fact that in many cases, the cells of interest may be in the minority and may be surrounded by normal or other abnormal cells, limits the ultimate utility of whole-tissue proteome studies in many instances.…”

Recent Developments in Capillary Electrophoresis–Mass Spectrometry of Proteins and Peptides

“…[67][68][69][70][71][72][73][74][75][76][77][78][79][80][81][82][83] Only these two configurations are covered in this review, although, brief descriptions of the other three are given in the succeeding paragraphs.…”

Progress in capillary electrophoresis coupled to matrix‐assisted laser desorption/ionization – time of flight mass spectrometry

“…A comprehensive 2-D micro-HPLC (m-HPLC) and CE system coupled with MALDI-TOF-TOF-MS analyzer was demonstrated by Zhang et al [23]. Protein/peptide samples were separated first with m-HPLC in RP mode with silica C8-packed capillary, and then the effluents were continuously transferred into CE through a novel valvefree hydrodynamic sampling interface.…”