TY - JOUR
T1 - Tracking the Stability of Clinically Relevant Blood Plasma Proteins with Delta-S-Cys-Albumin—A Dilute-and-Shoot LC/MS-Based Marker of Specimen Exposure to Thawed Conditions
AU - Kapuruge, Erandi P.
AU - Jehanathan, Nilojan
AU - Rogers, Stephen P.
AU - Williams, Stacy
AU - Chung, Yunro
AU - Borges, Chad R.
N1 - Funding Information:
here was supported in part by the National Cancer Institute of the National Institutes of Health under award number R33CA217702 (to C. R. B.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Funding Information:
We are grateful to Dr Stephanie M. F. Thibert, Dr Jesús Aguilar Díaz de león, and Ms Kazi Waheeda for their assistance with specimen processing and other time course logistics. We would also like to thank the NCI funded Cooperative Human Tissue Network (CHTN; Nashville, TN) and Valleywise Health (Phoenix, AZ) for providing biospecimens. Other investigators may have received aliquots from the same plasma/serum specimens provided by CHTN. The research reported here was supported in part by the National Cancer Institute of the National Institutes of Health under award number R33CA217702 (to C. R. B.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Publisher Copyright:
© 2022 THE AUTHORS. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology.
PY - 2022/11
Y1 - 2022/11
N2 - Biomolecular integrity can be compromised when blood plasma/serum (P/S) specimens are improperly handled. Compromised analytes can subsequently produce erroneous results—without any indication of having done so. We recently introduced an LC/MS-based marker of P/S exposure to thawed conditions called ΔS-Cys-Albumin which, aided by an established rate law, quantitatively tracks exposure of P/S to temperatures greater than their freezing point of −30 ◦C. The purposes of this study were to (1) evaluate ΔS-Cys-Albumin baseline values in gastrointestinal cancer patients and cancer-free control donors, (2) empirically assess the kinetic profiles of ΔS-Cys-Albumin at 23 ◦C, 4 ◦C, and −20 ◦C, and (3) empirically link ΔS-Cys-Albumin to the stability of clinically relevant proteins. ΔS-Cys-Albumin was measured at ≥ 9 different time points per exposure temperature in serum and K2EDTA plasma samples from 24 separate donors in aliquots kept separately at 23 ◦C, 4 ◦C, and −20 ◦C. Twenty-one clinically relevant plasma proteins were measured at four time points per temperature via a multiplexed immunoassay on the Luminex platform. Protein stability was assessed by mixed effects models. Coordinated shifts in stability between ΔS-Cys-Albumin and the unstable proteins were documented by repeated measures and Pearson correlations. Plasma ΔS-Cys-Albumin dropped from approximately 20% to under 5% within 96 h at 23 ◦C, 28 days at 4 ◦C, and 65 days at −20 ◦C. On average, 22% of the 21 proteins significantly changed in apparent concentration at each exposure temperature (p < 0.0008 with >10% shift). A linear inverse relationship was found between the percentage of proteins destabilized and ΔS-Cys-Albumin (r = −0.61; p < 0.0001)—regardless of the specific time/temperature of exposure. ΔS-Cys-Albumin tracks cumulative thawed-state exposure. These results now enable ΔS-Cys-Albumin to approximate the percentage of clinically relevant proteins that have been compromised by incidental plasma exposure to thawed-state conditions.
AB - Biomolecular integrity can be compromised when blood plasma/serum (P/S) specimens are improperly handled. Compromised analytes can subsequently produce erroneous results—without any indication of having done so. We recently introduced an LC/MS-based marker of P/S exposure to thawed conditions called ΔS-Cys-Albumin which, aided by an established rate law, quantitatively tracks exposure of P/S to temperatures greater than their freezing point of −30 ◦C. The purposes of this study were to (1) evaluate ΔS-Cys-Albumin baseline values in gastrointestinal cancer patients and cancer-free control donors, (2) empirically assess the kinetic profiles of ΔS-Cys-Albumin at 23 ◦C, 4 ◦C, and −20 ◦C, and (3) empirically link ΔS-Cys-Albumin to the stability of clinically relevant proteins. ΔS-Cys-Albumin was measured at ≥ 9 different time points per exposure temperature in serum and K2EDTA plasma samples from 24 separate donors in aliquots kept separately at 23 ◦C, 4 ◦C, and −20 ◦C. Twenty-one clinically relevant plasma proteins were measured at four time points per temperature via a multiplexed immunoassay on the Luminex platform. Protein stability was assessed by mixed effects models. Coordinated shifts in stability between ΔS-Cys-Albumin and the unstable proteins were documented by repeated measures and Pearson correlations. Plasma ΔS-Cys-Albumin dropped from approximately 20% to under 5% within 96 h at 23 ◦C, 28 days at 4 ◦C, and 65 days at −20 ◦C. On average, 22% of the 21 proteins significantly changed in apparent concentration at each exposure temperature (p < 0.0008 with >10% shift). A linear inverse relationship was found between the percentage of proteins destabilized and ΔS-Cys-Albumin (r = −0.61; p < 0.0001)—regardless of the specific time/temperature of exposure. ΔS-Cys-Albumin tracks cumulative thawed-state exposure. These results now enable ΔS-Cys-Albumin to approximate the percentage of clinically relevant proteins that have been compromised by incidental plasma exposure to thawed-state conditions.
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U2 - 10.1016/j.mcpro.2022.100420
DO - 10.1016/j.mcpro.2022.100420
M3 - Article
C2 - 36182099
AN - SCOPUS:85141658293
VL - 21
JO - Molecular and Cellular Proteomics
JF - Molecular and Cellular Proteomics
SN - 1535-9476
IS - 11
M1 - 100420
ER -