Elevated plasma albumin and apolipoprotein A-I oxidation under suboptimal specimen storage conditions

Chad Borges, Douglas S. Rehder, Sally Jensen, Matthew R. Schaab, Nisha D. Sherma, Hussein Yassine, Boriana Nikolova, Christian Breburda

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13 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1890-1899
Number of pages10
JournalMolecular and Cellular Proteomics
Volume13
Issue number7
DOIs
StatePublished - Jul 2014

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ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Molecular Biology

Cite this

Borges, C., Rehder, D. S., Jensen, S., Schaab, M. R., Sherma, N. D., Yassine, H., Nikolova, B., & Breburda, C. (2014). Elevated plasma albumin and apolipoprotein A-I oxidation under suboptimal specimen storage conditions. Molecular and Cellular Proteomics, 13(7), 1890-1899. https://doi.org/10.1074/mcp.M114.038455