Effects of gamma irradiation and 15 days of subsequent ex vivo storage on the cytosolic red blood cell proteome analyzed by 2D-DIGE and Orbitrap MS

Publication: Contribution to journalJournal articlesResearch

Abstract

PURPOSE: Gamma irradiation of red blood cell (RBC) concentrates is routinely used to prevent transfusion-associated graft-versus-host disease. So far, the effects of ionizing radiation on RBC structure and function and especially the proteome are not fully understood.

EXPERIMENTAL DESIGN: RBC concentrates were irradiated with 30 Gy and stored for 1 or 15 days at 4 ± 2°C. Following cell lysis and hemoglobin depletion, 2D-DIGE was used to examine the changes of the cytosolic RBC proteome. Significantly altered spots were analyzed using bottom-up proteomic approaches and selected marker proteins validated by western blotting.

RESULTS: Gamma irradiation was found to enhance conventional RBC storage lesions. Following 15 days of postirradiation storage, the abundances of a total of 27 spots were significantly altered and 3 out of 13 identified proteins were selected and validated as potential marker proteins for the assessment of irradiation-induced cytosolic RBC lesions.

CONCLUSIONS AND CLINICAL RELEVANCE: Gamma irradiation and subsequent ex vivo storage according to the Council of Europe guidelines were found to affect RBC protein structures. The validated marker proteins can serve as a basis for the development of a screening assay to monitor the quality of irradiated RBC concentrates during ex vivo storage.

Original languageEnglish
JournalProteomics. Clinical applications
Volume7
Issue number7-8
Pages (from-to)561-570
Number of pages10
DOIs
Publication statusPublished - 01.08.2013

Research areas and keywords

  • Adult
  • Biological Markers
  • Chromatography, Liquid
  • Cytosol
  • Erythrocytes
  • Female
  • Gamma Rays
  • Humans
  • Male
  • Mass Spectrometry
  • Nanotechnology
  • Proteomics
  • Specimen Handling
  • Time Factors
  • Two-Dimensional Difference Gel Electrophoresis
  • Young Adult

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