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Proteomic analysis of red blood cells from donors exhibiting high hemolysis demonstrates a reduction in membrane‐associated proteins involved in the oxidative response
Author(s) -
Chen Deborah,
Schubert Peter,
Devine Dana V.
Publication year - 2017
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/trf.14188
Subject(s) - hemolysis , red blood cell , erythrocyte fragility , hemoglobin , ex vivo , quantitative proteomics , andrology , microvesicle , proteomics , mean corpuscular hemoglobin , chemistry , biology , immunology , biochemistry , medicine , mean corpuscular volume , microvesicles , in vitro , microrna , gene
BACKGROUND The development of hemolysis during ex vivo hypothermic storage is multifaceted. Standardization of collection and production processes is used to minimize variability in biologics manufacturing and to maximize product quality. However, the influence of various donor characteristics on product quality is often difficult to evaluate and to control. Using a proteomic approach, we aimed to decipher relevant donor characteristics that may predict red blood cell (RBC) quality during storage. STUDY DESIGN AND METHODS Ten healthy volunteer donors exhibiting repeated high hemolysis at outdate (>0.8%; RBC HH ) and 10 age‐ and sex‐matched control donors (RBC Ctrl ) were studied. Common quality variables were measured on Days 5, 14, 21, 28, and 42 of storage. Protein profiles of hemoglobin‐depleted membrane fractions from RBC HH and RBC Ctrl donors were analyzed using a quantitative proteomics approach based on iTRAQ (isobaric tags for relative and absolute quantitation). RESULTS Time‐dependent lesion development was apparent in both donor populations. RBC HH exhibited reduced 2,3‐bisphosphoglycerate levels (p < 0.001) and morphologic score (p < 0.001), but displayed elevated hemolysis level (p < 0.001), RBC‐derived microvesicle formation (p < 0.001), and mean corpuscular fragility (p < 0.001) compared to RBC Ctrl , indicating notable differences at the membrane between the two donor populations. Proteomic findings revealed a significant reduction in the level of proteins involved in oxidative response pathways at early time points in RBC HH compared to that of RBC Ctrl . CONCLUSION The recruitment of these candidate proteins might be part of a response mechanism altered in RBC HH donors and therefore may be useful as a donor screening tool.