Exposure of blood plasma/serum (P/S) to thawed conditions (> -30 °C) can produce biomolecular changes that skew measurements of biomarkers within archived patient samples, potentially rendering them unfit for molecular analysis. Because freeze-thaw histories are often poorly documented, objective methods for assessing molecular fitness before analysis are needed. We report a 10-μl, dilute-and-shoot, intact-protein mass spectrometric assay of albumin proteoforms called “Δ S-Cys-Albumin” that quantifies cumulative exposure of archived P/S samples to thawed conditions. The relative abundance of S-cysteinylated (oxidized) albumin in P/S increases inexorably but to a maximum value under 100% when samples are exposed to temperatures > -30 °C. The difference in the relative abundance of S-cysteinylated albumin (S-Cys-Alb) before and after an intentional incubation period that drives this proteoform to its maximum level is denoted as Δ S-Cys-Albumin. Δ S-Cys-Albumin in fully expired samples is zero. The range (mean ± 95% CI) observed for Δ S-Cys-Albumin in fresh cardiac patient P/S (n = 97) was, for plasma 12-29% (20.9 ± 0.75%) and for serum 10 -24% (15.5 ± 0.64%). The multireaction rate law that governs S-Cys-Alb formation in P/S was determined and shown to predict the rate of formation of S-Cys-Alb in plasma and serum samples-a step that enables back-calculation of the time at which unknown P/S specimens have been exposed to room temperature. A blind challenge demonstrated that Δ S-Cys-Albumin can detect exposure of groups (n = 6 each) of P/S samples to 23 °C for 2 h, 4 °C for 16 h, or -20 °C for 24 h-and exposure of individual specimens for modestly increased times. An unplanned case study of nominally pristine serum samples collected under NIH-sponsorship demonstrated that empirical evidence is required to ensure accurate knowledge of archived P/S biospecimen storage history.
CITATION STYLE
Jeffs, J. W., Jehanathan, N., Thibert, S. M. F., Ferdosi, S., Pham, L., Wilson, Z. T., … Borges, C. R. (2019). Delta-S-cys-albumin: A lab test that quantifies cumulative exposure of archived human blood plasma and serum samples to thawed conditions. Molecular and Cellular Proteomics, 18(10), 2121–2137. https://doi.org/10.1074/mcp.TIR119.001659
Mendeley helps you to discover research relevant for your work.