title: Elevated plasma albumin and apolipoprotein A-I oxidation under suboptimal specimen storage conditions.
abstract: S-cysteinylated albumin and methionine-oxidized apolipoprotein A-I (apoA-I) have been posed as candidate markers of diseases associated with oxidative stress. Here, a dilute-and-shoot form of LC-electrospray ionization-MS requiring half a microliter of blood plasma was employed to simultaneously quantify the relative abundance of these oxidized proteoforms in samples stored at -80 °C, -20 °C, and room temperature and exposed to multiple freeze-thaw cycles and other adverse conditions in order to assess the possibility that protein oxidation may occur as a result of poor sample storage or handling. Samples from a healthy donor and a participant with poorly controlled type 2 diabetes started at the same low level of protein oxidation and behaved similarly; significant increases in albumin oxidation via S-cysteinylation were found to occur within hours at room temperature and days at -20 °C. Methionine oxidation of apoA-I took place on a longer time scale, setting in after albumin oxidation reached a plateau. Freeze-thaw cycles had a minimal effect on protein oxidation. In matched collections, protein oxidation in serum was the same as that in plasma. Albumin and apoA-I oxidation were not affected by sample headspace or the degree to which vials were sealed. ApoA-I, however, was unexpectedly found to oxidize faster in samples with lower surface-area-to-volume ratios. An initial survey of samples from patients with inflammatory conditions normally associated with elevated oxidative stress-including acute myocardial infarction and prostate cancer-demonstrated a lack of detectable apoA-I oxidation. Albumin S-cysteinylation in these samples was consistent with known but relatively brief exposures to temperatures above -30 °C (the freezing point of blood plasma). Given their properties and ease of analysis, these oxidized proteoforms, once fully validated, may represent the first markers of blood plasma specimen integrity based on direct measurement of oxidative molecular damage that can occur under suboptimal storage conditions.
authors.affiliation: From the ‡Department of Chemistry & Biochemistry, Arizona State University, Tempe, Arizona 85287; §Biodesign Institute at Arizona State University, Tempe, Arizona 85287; chad.borges@asu.edu.
authors.forename: Chad R
authors.initials: CR
authors.lastname: Borges
authors.affiliation: §Biodesign Institute at Arizona State University, Tempe, Arizona 85287;
authors.forename: Douglas S
authors.initials: DS
authors.lastname: Rehder
authors.affiliation: From the ‡Department of Chemistry & Biochemistry, Arizona State University, Tempe, Arizona 85287;
authors.forename: Sally
authors.initials: S
authors.lastname: Jensen
authors.affiliation: §Biodesign Institute at Arizona State University, Tempe, Arizona 85287;
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Schaab
authors.affiliation: §Biodesign Institute at Arizona State University, Tempe, Arizona 85287;
authors.forename: Nisha D
authors.initials: ND
authors.lastname: Sherma
authors.affiliation: ‖Department of Medicine, University of Southern California, Los Angeles, California 90033;
authors.forename: Hussein
authors.initials: H
authors.lastname: Yassine
authors.affiliation: **Maricopa Integrated Health Systems, Phoenix, Arizona 85008;
authors.forename: Boriana
authors.initials: B
authors.lastname: Nikolova
authors.affiliation: **Maricopa Integrated Health Systems, Phoenix, Arizona 85008; ‡‡College of Medicine, University of Arizona, Phoenix, Arizona 85004.
authors.forename: Christian
authors.initials: C
authors.lastname: Breburda
mesh_terms: D016632:Apolipoprotein A-I / Q000032:analysis* / Q000097:blood / Q000378:metabolism; D002853:Chromatography, Liquid; D015925:Cryopreservation / Q000379:methods; D003545:Cysteine / Q000737:chemistry*; D003924:Diabetes Mellitus, Type 2; D005615:Freezing; D006801:Humans; D008297:Male; D008715:Methionine / Q000737:chemistry*; D009203:Myocardial Infarction; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011471:Prostatic Neoplasms; D012709:Serum Albumin / Q000032:analysis* / Q000378:metabolism; D013048:Specimen Handling; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D016632:Apolipoprotein A-I; D012709:Serum Albumin; D008715:Methionine; D003545:Cysteine
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