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Complex proteome prefractionation using microscale solution isoelectrofocusing
Abstract:The proteomes of mammalian cells, tissues and biologic fluids are complex and consist of proteins present over a wide dynamic range. Current protein profiling technologies do not have the capacity to overcome the sample complexity for comprehensive analysis of complex proteomes. A common strategy to substantially expand protein profiling capacities is sample prefractionation. A prefractionation method developed in the authors’ laboratory, microscale solution isoelectrofocusing, has resulted in a commercial product, the ZOOM® IEF Fractionator, which provides a simple and convenient method for high-resolution separation of complex proteomes based upon their isoelectric points. Complex human samples such as cancer cells and biologic fluids can be fractionated into well-resolved fractions with minimal cross-contamination of proteins between adjacent fractions. This review focuses on the application of microscale solution isoelectrofocusing prefractionation and subsequent downstream strategies in expanding protein profiling capacities and mining low-abundance proteins of complex proteomes.
Keywords:2D arrays  liquid chromatography tandem mass spectrometry  microscale solution isoelectrofocusing  proteomics methods  sample prefractionation  ZOOM® IEF
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