On-chip microextraction for proteomic sample preparation of in-gel digests

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Bibtex

@article{3d964cccc2d84673977529ee38ea3ac4,
title = "On-chip microextraction for proteomic sample preparation of in-gel digests",
abstract = "Despite the high sensitivity and relatively high tolerance for contaminants of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) there is often a need to purify and concentrate the sample solution, especially after in-gel digestion of proteins separated by two-dimensional gel electrophoresis (2-DE). A silicon microextraction chip (SMEC) for sample clean-up and trace enrichment of peptides was manufactured and investigated. The microchip structure was used to trap reversed-phase chromatography media (POROS R2 beads) that facilitates sample purification/enrichment of contaminated and dilute samples prior to the MALDI-TOF MS analysis. The validity of the SMEC sample preparation technique was successfully investigated by performing analysis on a 10 nM peptide mixture containing 2 m urea in 0.1 m phosphate-buffered saline with MALDI-TOF MS. It is demonstrated that the microchip sample clean-up and enrichment of peptides can facilitate identification of proteins from 2-DE separations. The microchip structure was also used to trap beads immobilized with trypsin, thereby effectively becoming a microreactor for enzymatic digestion of proteins. This microreactor was used to generate a peptide map from a 100 nM bovine serum albumin sample.",
keywords = "Electrophoresis, Gel, Two-Dimensional, Enzymes, Immobilized, Peptides, Protein Array Analysis, Proteome, Reproducibility of Results, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Evaluation Studies, Journal Article, Research Support, Non-U.S. Gov't",
author = "Simon Ekstr{\"o}m and Johan Malmstr{\"o}m and Lars Wallman and Mikael L{\"o}fgren and Johan Nilsson and Thomas Laurell and Gy{\"o}rgy Marko-Varga",
year = "2002",
month = "4",
doi = "10.1002/1615-9861(200204)2:4<413::AID-PROT413>3.0.CO;2-1",
language = "English",
volume = "2",
pages = "413--421",
journal = "Proteomics",
issn = "1615-9861",
publisher = "John Wiley & Sons",
number = "4",

}