Peroxiredoxin-2 as a candidate biomarker to test oxidative stress levels of stored red blood cells under blood bank conditions.

Author(s): Rinalducci S, D'Amici GM, Blasi B, Vaglio S, Grazzini G, Zolla L

Publication: Transfusion, 2011, Vol. 51, Page 1439-49

PubMed ID: 21276001 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of refrigerated storage of RBCs under aerobic and anaerobic conditions on membrane-bound protein content. Blood was collected in citrate-phosphate-dextrose (CPD), and leukoreduced RBCs were suspended in saline-adenine-glucose-mannitol (SAGM). Specimens were centrifuged and transferred to new polyvinylchloride bags, additive solution (AS) was added to each bag, and helium was used for oxygen depletion. After storage, RBC membrane proteins were extracted, analyzed by electrophoresis and immunoblotting, and specific bands were cut from gels for protein identification by tandem mass spectrometry.

   Summary of Findings:

   During storage, the density of a high molecular band (>205 kDa) identified as an Hb-cytoskeletal protein cross-link increased under reducing and non-reducing conditions. The density of the CAT band in reducing gels increased with increasing storage duration, but the band was not visible in non-reducing gels or after anaerobic storage. The band corresponding to 2,3 BGM appeared in specimens stored for 42 days, regardless of gel type or the presence of oxygen. Prx2 increased with storage, first becoming evident after 21 days of storage, and in non-reducing conditions, was present as a dimer. Prx2 was less abundant after anaerobic storage than aerobic storage. Finally, storage caused an increase in membrane bound Hb dimers. Between specimens the timing and level of increase in Prx2 was variable, with first appearance occurring between day 14 and 25. MDA increased with increasing storage duration, regardless of donor.

   Biospecimens

   • Cell - Red Blood Cells

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Protein	1D/2D gels
   Protein	Western blot
   Protein	LC-MS or LC-MS/MS
   Peptide	LC-MS or LC-MS/MS
   Small molecule	Spectrophotometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage duration	0 days 7 days 14 days 21 days 28 days 42 days
   Western blot Specific	Targeted peptide/protein	Prx2
   1D/2D gels Specific	Technology platform	Reducing gel Non-reducing gel
   Storage	Storage conditions	Aerobic Anaerobic

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