title: Combination of affinity depletion of abundant proteins and reversed-phase fractionation in proteomic analysis of human plasma/serum.
abstract: The tremendous complexity of the serum and plasma proteome presents extreme analytical challenges in comprehensive analysis due to the wide dynamic range of protein concentrations. Therefore, robust sample preparation methods remain one of the important steps in the proteome characterization workflow. We present the results on a new column for the specific depletion of 14 high-abundant proteins from human serum and plasma and the subsequent reversed-phase fractionation of the flow-through proteins. Analysis of tryptic peptides was accomplished with microfluidic HPLC-Chip/MS system. Results indicate that high-abundant protein depletion combined with RP fractionation of plasma showed an improved dynamic range for proteomic analysis and enabled the identification of low-abundant plasma proteins.
authors.affiliation: Agilent Technologies Inc., 2850 Centerville Road, Wilmington, DE 19808, USA. nina_zolotarjova@agilent.com
authors.forename: Nina
authors.initials: N
authors.lastname: Zolotarjova
authors.forename: Peter
authors.initials: P
authors.lastname: Mrozinski
authors.forename: Haiying
authors.initials: H
authors.lastname: Chen
authors.forename: James
authors.initials: J
authors.lastname: Martosella
mesh_terms: D001798:Blood Proteins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D005591:Chemical Fractionation; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D020543:Proteome / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D001798:Blood Proteins; D020543:Proteome