title: Highly sensitive proteome analysis of FACS-sorted adult colon stem cells.
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.
authors.affiliation: Biomolecular Mass Spectrometry and Proteomics Group, Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Serena
authors.initials: S
authors.lastname: Di Palma
authors.forename: Daniel
authors.initials: D
authors.lastname: Stange
authors.forename: Marc
authors.initials: M
authors.lastname: van de Wetering
authors.forename: Hans
authors.initials: H
authors.lastname: Clevers
authors.forename: Albert J R
authors.initials: AJ
authors.lastname: Heck
authors.forename: Shabaz
authors.initials: S
authors.lastname: Mohammed
mesh_terms: D000328:Adult; D002469:Cell Separation / Q000379:methods*; D002853:Chromatography, Liquid; D003106:Colon / Q000166:cytology / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry; D020543:Proteome; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*
chemical_list: D020543:Proteome