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Free‐flow electrophoresis in proteome sample preparation
Author(s) -
Wildgruber Robert,
Weber Gerhard,
Wise Petra,
Grimm Daniela,
Bauer Johann
Publication year - 2014
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201300253
Subject(s) - isotachophoresis , free flow electrophoresis , proteome , proteomics , electrophoresis , chromatography , two dimensional gel electrophoresis , capillary electrophoresis , chemistry , gel electrophoresis , biology , organelle , peptide , biochemistry , gel electrophoresis of proteins , polyacrylamide gel electrophoresis , enzyme , gene , electrode , electrolyte
An aim of proteome research is to identify the entire complement of proteins expressed in defined cell types of humans, animals, plants, and microorganisms. The approach requires searching for low abundant or even rarely expressed proteins in many cell types, as well as the determination of the protein expression levels in subcellular compartments and organelles. In recent years, rather powerful MS technologies have been developed. At this stage of MS device development, it is of highest interest to purify intact cell types or isolate subcellular compartments, where the proteins of interest are originating from, which determine the final composition of a peptide mixture. Free‐flow electrophoresis proved to be useful to prepare meaningful peptide mixtures because of its improved capabilities in particle electrophoresis and the enhanced resolution in protein separation. Sample preparation by free‐flow electrophoresis mediated particle separation was preferentially performed for purification of either organelles and their subspecies or major protein complexes. Especially, the introduction of isotachophoresis and interval zone electrophoresis improved the purity of the gained analytes of interest. In addition, free‐flow IEF proved to be helpful, when proteins of low solubility, obtained, e.g. from cell membranes, were investigated.