title: 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.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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
  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.
authors.affiliation: Center for Preventive Doping Research/Institute of Biochemistry, German Sport University Cologne, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Walpurgis
authors.forename: Maxie
authors.initials: M
authors.lastname: Kohler
authors.forename: Andreas
authors.initials: A
authors.lastname: Thomas
authors.forename: Folker
authors.initials: F
authors.lastname: Wenzel
authors.forename: Hans
authors.initials: H
authors.lastname: Geyer
authors.forename: Wilhelm
authors.initials: W
authors.lastname: Schänzer
authors.forename: Mario
authors.initials: M
authors.lastname: Thevis
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000097:blood; D002853:Chromatography, Liquid; D003600:Cytosol / Q000378:metabolism* / Q000528:radiation effects*; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism* / Q000528:radiation effects*; D005260:Female; D005720:Gamma Rays / Q000009:adverse effects*; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D036103:Nanotechnology; D040901:Proteomics / Q000379:methods*; D013048:Specimen Handling / Q000379:methods*; D013997:Time Factors; D057825:Two-Dimensional Difference Gel Electrophoresis; D055815:Young Adult
chemical_list: D015415:Biomarkers
keywords: 2D-DIGE; Gamma irradiation; Red blood cells; Storage lesions; Transfusion medicine