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Highly sensitive proteome analysis of FACS-sorted adult colon stem cells.

Research paper by Serena S Di Palma, Daniel D Stange, Marc M van de Wetering, Hans H Clevers, Albert J R AJ Heck, Shabaz S Mohammed

Indexed on: 24 Jun '11Published on: 24 Jun '11Published in: Journal of Proteome Research



Abstract

In proteomics, multidimensional liquid chromatography combined with mass spectrometry has become a standard technique to reduce sample complexity and tackle the vast dynamic range. Such fractionation is necessary to obtain a comprehensive analysis of biological samples such as tissues and cell lines. However, extensive fractionation comes at the expense of sample losses, hampering the analysis of limited material. We previously described a highly sensitive multidimensional chromatographic strategy based on a combination of hydrophilic interaction liquid chromatography and reversed phase chromatography, which allows proteomic analysis with minimal sample losses. Here we apply this strategy to the analysis of a limited number of FACS-sorted colon stem cells extracted from mouse intestine, obtaining a proteome coverage comparable to current methods that generally require 100-fold more starting material. We propose that this alternative multidimensional chromatographic technology will find ample application such as in the analysis of distinct cellular populations obtained by laser microdissection.