Proteomic analysis of plasma derived from platelet buffy coats during storage at room temperature. An application of ProteoMiner technology.
Author(s): Egidi MG, Rinalducci S, Marrocco C, Vaglio S, Zolla L
Publication: Platelets, 2011, Vol. 22, Page 252-69
PubMed ID: 21405958 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to determine the effects of storage at room temperature and protein equalization on the proteomic profile of platelet concentrate (PC) supernatants.
Conclusion of Paper
Use of ProteoMiner increased the number of spots reproducibly identified by 2-D gels of PC supernatant. ProteoMiner allowed for detection of 45 spots that were affected by storage of platelet buffy coats compared to 12 when supernatant was not treated.
Studies
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Study Purpose
The purpose of this study was to determine the effects of storage at room temperature and protein equalization on the proteomic profile of PC supernatants. Citrate phosphate dextrose (CPD)- saline, adenine, glucose and manitol (SAGM) blood was processed to buffy coat after <6 h at room temperature, and PC was obtained from buffy coat after 1-2 h at room temperature. PC was stored under standard room temperature conditions with agitation, and supernatant was sampled aseptically at each time point after soft spin centrifugation. Supernatants were frozen before proteomic analysis.
Summary of Findings:
Use of ProteoMiner to reduce variability in protein concentrations allowed for detection of 20% more spots in 2D gels and decreased the intensity of spots of high concentration proteins like albumin. 570 spots were reproducibly identified across all 2D gels of untreated supernatant, of which 6 spots decreased and 6 spots increased by more than 2-fold during storage (p<0.05). The increases in kininogen 1 (KNG1) and SERPINA8 proteins were confirmed by Western blot (only two proteins examined). 2D gels of ProteoMiner treated supernatant had 717 spots with reliable detection across all gels, with 13 spots increasing, and 26 decreasing more than 2-fold between day 0 and 7 (p<0.05). An additional 3 spots increased between day 0 and 4 (p<0.05) but then decreased between day 4 and 7 (p<0.05), and 1 spot decreased between day 0 and 4 (p<0.05) and then increased between day 4 and 7 (p<0.05). The authors conclude that ProteoMiner allowed for detection of previously undetected proteins in PC supernatant that are affected by storage.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Normal
Platform:
Analyte Technology Platform Protein 1D/2D gels Protein MS/MS Protein Western blot Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage duration 0 days
4 days
7 days
1D/2D gels Specific Signal amplification None
ProteoMiner