title: Integrated capillary isoelectric focusing/nano-reversed phase liquid chromatography coupled with ESI-MS for characterization of intact yeast proteins.
abstract: An integrated protein concentration/separation platform, combining capillary isoelectric focusing (CIEF) with nano-reversed phase liquid chromatography (nano-RPLC), is developed to provide significant protein concentration and high resolving power for the analysis of complex protein mixtures. Upon completion of protein focusing, the proteins are sequentially and hydrodynamically loaded into individual trap columns using a group of microinjection and microselection valves. Repeated pro-tein loadings and injections into trap columns are carried out automatically until the entire CIEF cap-illary content is sampled and fractionated. Each CIEF fraction "parked" in separate trap columns is further resolved using nano-RPLC, and the eluants are analyzed using electrospray ionization-mass spectrometry.
authors.affiliation: Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA.
authors.forename: Yueju
authors.initials: Y
authors.lastname: Wang
authors.forename: Brian M
authors.initials: BM
authors.lastname: Balgley
authors.forename: Paul A
authors.initials: PA
authors.lastname: Rudnick
authors.forename: Erin L
authors.initials: EL
authors.lastname: Evans
authors.forename: Don L
authors.initials: DL
authors.lastname: DeVoe
authors.forename: Cheng S
authors.initials: CS
authors.lastname: Lee
mesh_terms: D001331:Automation; D002458:Cell Fractionation; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D005656:Fungal Proteins / Q000032:analysis / Q000302:isolation & purification*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D012995:Solubility; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation*
chemical_list: D005656:Fungal Proteins