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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
Author(s) -
Walpurgis Katja,
Kohler Maxie,
Thomas Andreas,
Wenzel Folker,
Geyer Hans,
Schänzer Wilhelm,
Thevis Mario
Publication year - 2013
Publication title -
proteomics – clinical applications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.948
H-Index - 54
eISSN - 1862-8354
pISSN - 1862-8346
DOI - 10.1002/prca.201300009
Subject(s) - ex vivo , proteome , hemoglobin , in vivo , irradiation , lysis , red blood cell , cytosol , chemistry , biology , microbiology and biotechnology , andrology , biochemistry , enzyme , medicine , genetics , physics , nuclear physics
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.