title: Assessment of albumin removal from an immunoaffinity spin column: critical implications for proteomic examination of the albuminome and albumin-depleted samples.
abstract: High abundance proteins in serum and plasma (e.g., albumin) are routinely removed during proteomic sample processing as they can mask lower abundance proteins and peptides of biological/clinical interest. A common method of albumin depletion is based on immunoaffinity capture, and many immunoaffinity devices are designed for multiple uses. In this case, it is critical that the albumin captured on the affinity matrix is stripped from the column prior to regeneration of the matrix and processing of subsequent samples, to ensure no carryover and that maximal binding sites are available for subsequent samples. The current study examines the ability of a manufacturer's protocol to remove the proteins and peptides captured by an immunoaffinity spin column. The data presented in the current work illustrate the difficulty in completely removing albumin from the immunoaffinity device, and consequently, may explain the variability and decreased efficiency shown for this device in previous studies. In summary, the current data present important considerations for the implementation of multiple-use immunoaffinity devices for processing subsequent clinical samples in a proteomic workflow.
authors.affiliation: Department of Medicine, Johns Hopkins University, Baltimore, MD21224, USA.
authors.forename: Rebekah L
authors.initials: RL
authors.lastname: Gundry
authors.forename: Melanie Y
authors.initials: MY
authors.lastname: White
authors.forename: Julie
authors.initials: J
authors.lastname: Nogee
authors.forename: Irina
authors.initials: I
authors.lastname: Tchernyshyov
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Van Eyk
mesh_terms: D002846:Chromatography, Affinity / Q000295:instrumentation*; D002851:Chromatography, High Pressure Liquid; D017743:Equipment Reuse; D006801:Humans; D007120:Immunochemistry / Q000379:methods*; D040901:Proteomics; D012709:Serum Albumin / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D012709:Serum Albumin