title: A fast and reproducible method for albumin isolation and depletion from serum and cerebrospinal fluid.
abstract: Analysis of serum and plasma proteomes is a common approach for biomarker discovery, and the removal of high-abundant proteins, such as albumin and immunoglobins, is usually the first step in the analysis. However, albumin binds peptides and proteins, which raises concerns as to how the removal of albumin could impact the outcome of the biomarker study while ignoring the possibility that this could be a biomarker subproteome itself. The first goal of this study was to test a new commercially available affinity capture reagent from Protea Biosciences and to compare the efficiency and reproducibility to four other commercially available albumin depletion methods. The second goal of this study was to determine if there is a highly efficient albumin depletion/isolation system that minimizes sample handling and would be suitable for large numbers of samples. Two of the methods tested (Sigma and ProteaPrep) showed an albumin depletion efficiency of 97% or greater for both serum and cerebrospinal fluid (CSF). Isolated serum and CSF albuminomes from ProteaPrep spin columns were analyzed directly by LC-MS/MS, identifying 128 serum (45 not previously reported) and 94 CSF albuminome proteins (17 unique to the CSF albuminome). Serum albuminome was also isolated using Vivapure anti-HSA columns for comparison, identifying 105 proteins, 81 of which overlapped with the ProteaPrep method.
authors.affiliation: Department of Medicine, Division of Cardiology, Johns Hopkins University, Baltimore, MD 21224, USA.
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Holewinski
authors.forename: Zhicheng
authors.initials: Z
authors.lastname: Jin
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Powell
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Maust
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Van Eyk
mesh_terms: D000418:Albumins / Q000032:analysis / Q000134:cerebrospinal fluid* / Q000737:chemistry / Q000302:isolation & purification*; D015415:Biomarkers / Q000097:blood / Q000134:cerebrospinal fluid / Q000737:chemistry; D015153:Blotting, Western; D002853:Chromatography, Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D020543:Proteome / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000418:Albumins; D015415:Biomarkers; D020543:Proteome