title: Free-flow electrophoresis of the human urinary proteome.
abstract: Prefractionation of complex protein samples prior to mass spectrometry provides a method for the isolation of low-abundance proteins into specific fractions thereby enabling their identification. Free-flow electrophoresis in the isoelectric focusing mode (IEF-FFE) presents a complementary approach to established prefractionation methodologies. Proteins are separated in solution according to their isoelectric point (pI) with a high throughput of sample volume. The separation may be performed under denaturing or nondenaturing conditions and detergents may be added to promote protein solubilization. A protocol covering the pH range from pH 3 to 9 under denaturing conditions was used to illustrate the method of IEF-FFE including sample preparation prior to reversed-phase liquid chromatography and tandem mass spectrometry. The IEF-FFE separation was applied to a sample of human urine.
authors.affiliation: BD Diagnostics, Martinsried, Germany.
authors.forename: Mikkel
authors.initials: M
authors.lastname: Nissum
authors.forename: Robert
authors.initials: R
authors.lastname: Wildgruber
mesh_terms: D004586:Electrophoresis / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D007525:Isoelectric Focusing / Q000295:instrumentation / Q000379:methods; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D014556:Urine / Q000737:chemistry*
chemical_list: D020543:Proteome