Biochemically Tracked Variability of Blood Plasma Thawed-State Exposure Times in a Multisite Collection Study.
Author(s): Hu Y, Mulot C, Bourreau C, Martin D, Laurent-Puig P, Radoï L, Guénel P, Borges CR
Publication: Biopreserv Biobank, 2020, Vol. 18, Page 376-388
PubMed ID: 32608993 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to compare the relative abundance of S-cysteinylated albumin (𝛥S-Cys-Albumin) in plasma obtained after different processing delays or subjected to temporary storage at -20°C. The 𝛥S-Cys-Albumin values were also compared in plasma specimens from cancer patients and age-matched controls.
Conclusion of Paper
Pre-centrifugation delay and total processing delay were modestly correlated with 𝛥S-Cys-Albumin values with significantly higher 𝛥S-Cys-Albumin values in specimens with a prec-entrifugation or total processing delay of <17h. While 94% of specimens stored at -80°C within 3 h of blood draw had 𝛥S-Cys-Albumin values in the expected range (12-29%), 80% of specimens with a delay to frozen storage of 3-13 h and 54% of specimens with a delay >17 h had 𝛥S-Cys-Albumin values in the expected range. None of the specimens stored at -20°C before shipment had 𝛥S-Cys-Albumin values in the expected range. The values of 𝛥S-Cys-Albumin were dependent on collection site but the two sites that clustered most distinctly were the only ones that stored specimens at -20°C. S-Cys-Albumin and 𝛥S-Cys-Albumin values were significantly different in cancer and control patients with specimen from cancer patients having lower S-Cys-Albumin and higher 𝛥S-Cys-Albumin than controls but these differences were small compared to those attributed to processing/storage delays.
Studies
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Study Purpose
The purpose of this study was to compare the relative abundance of 𝛥S-Cys-Albumin in plasma obtained after different processing delays or subjected to temporary storage at -20°C. The 𝛥S-Cys-Albumin values were also compared in plasma specimens from cancer patients and age-matched controls. EDTA blood was collected from 206 Stage 1 and 4 lung cancer patients and 207 age/gender matched healthy controls at 12 different sites and transported to the laboratory on ice packs. Plasma was obtained by centrifugation at 3000 rpm for 15 min at 4°C within 45 min of blood collection. Plasma was aliquoted and frozen at -20°C (two sites) or -80°C until shipment to a central laboratory and storage at -80°C. Albumin levels were quantified by LC-ESI-MS and 𝛥S-Cys-Albumin values were calculated based on the relative abundance of S-Cys-Albumin. Only highly hemolyzed specimens (>250 mg hemoglobin/dL) were excluded from analysis. Specimens were categorized based on the time from draw until storage at -80°C (pre- and post-centrifugation combined) which was <3 h (124 specimens), 3-13 h (162 specimens), or >17 h (70 specimens) at room temperature or by storage of plasma at -20°C for 42-456 days (46 specimens) before shipment. The effect of pre-centrifugation delay (0-48 h) was investigated in specimens with a post-centrifugation delay to frozen storage at -80°C of <1.5 h. Due to the high level of variability in storage durations and temperatures, the authors used a model to correct 𝛥S-Cys-Albumin values to be the equivalent of storage at 23°C.
Summary of Findings:
𝛥S-Cys-Albumin levels decreased with increasing time from blood draw to storage at -80°C. Pre-centrifugation delay and total processing delay were modestly correlated with 𝛥S-Cys-Albumin (r=0.522, P<0.0001 and r=0.524, P<0.0001, respectively) with significantly higher 𝛥S-Cys-Albumin values in specimens with a pre-centrifugation or total processing delay of <17 h (P<0.00001, both). While 94% of specimens stored at -80°C within 3 h of blood draw had 𝛥S-Cys-Albumin values in the expected range (12-29%), 80% of specimens with a delay to frozen storage of 3-13 h and 54% of specimens with a delay to frozen storage >17 h had 𝛥S-Cys-Albumin values in the expected range. Further, none of the specimens stored at -20°C before shipment had 𝛥S-Cys-Albumin values in the expected range. The 𝛥S-Cys-Albumin values were dependent on collection site but the two sites that clustered most distinctly were the only ones that stored specimens at -20°C. Specimens stored at -20°C for exhibited lower values of 𝛥S-Cys-Albumin values reflecting an equivalent longer exposure to 23°C. S-Cys-Albumin and 𝛥S-Cys-Albumin values were significantly different in cancer and control patients with specimens from cancer patients having lower S-Cys-Albumin (30 ± 11 versus 32±7.2, P<0.01) and higher 𝛥S-Cys-Albumin (16±7.9 versus 14±5.6, P<0.01) than controls but these differences were small compared to those attributed to processing/storage delays.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Not specified
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform Protein LC-ESI-MS/MS Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Preaquisition Diagnosis/ patient condition Lung cancer patient
Control patient
Biospecimen Aliquots and Components Centrifugation Centrifugation delays investigated
Biospecimen Acquisition Locale of biospecimen collection 12 different collection sites compared
Storage Storage temperature -20°C
-80°C
Storage Time at room temperature 0-48 h
<3 h
3-13 h
<17 h