Supernatant protein biomarkers of red blood cell storage hemolysis as determined through an absolute quantification proteomics technology

Abstract

BACKGROUND Laboratory technologies have highlighted the progressive accumulation of the so-called “storage lesion,” a wide series of alterations to stored red blood cells (RBCs) that may affect the safety and effectiveness of the transfusion therapy. New improvements in the field are awaited to ameliorate this lesion, such as the introduction of washing technologies in the cell processing pipeline. Laboratory studies that have tested such technologies so far rely on observational qualitative or semiquantitative techniques. STUDY DESIGN AND METHODS A state-of-the-art quantitative proteomics approach utilizing quantitative concatamers (QconCAT) was used to simultaneously monitor fluctuations in the abundance of 114 proteins in AS-3 RBC supernatants (n =5; 11 time points, including before and after leukoreduction, at 3 hours, on Days 1 and 2, and weekly sampling from Day 7 through Day 42). RESULTS Leukoreduction-dependent depletion of plasma proteins was observed at the earliest time points. A subset of proteins showed very high linear correlation (r2 >0.9) not only with storage time, but also with absolute levels of hemoglobin α1 and β, a proxy for RBC hemolysis and vesiculation. Linear regression was performed to describe the temporal relationship between these proteins. Our findings suggest a role for supernatant glyceraldehyde-3-phosphate dehydrogenase; peroxiredoxin-1, -2, and -6; carbonic anhydrase-1 and -2; selenium binding protein-1; biliverdin reductase; aminolevulinate dehydratase; and catalase as potential biomarkers of RBC quality during storage. CONCLUSION A targeted proteomics technology revealed novel biomarkers of the RBC storage lesion and promises to become a key analytical readout for the development and testing of alternative cell processing strategies.

Freeze‐dried plasma enhances clot formation and inhibits fibrinolysis in the presence of tissue plasminogen activator similar to pooled liquid plasma

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Donor‐variation effect on red blood cell storage lesion: A close relationship emerges

“…RBCs undergo time‐dependent deterioration in several aspects of their physiology during storage in standard blood bank conditions, collectively named as “RBC storage lesion.” In this context, functionally important disturbances in energy and redox metabolism, rheology, and finally, in RBC aging and removal signaling are apparent after examination of RBC concentrates in vitro by both targeted and integrated, omics‐based approaches . Indeed, time‐dependent changes in the expression/accumulation and posttranslational modifications of hemoglobin (Hb), supernatant and membrane proteins and lipids, including fragmentation, oxidation/carbonylation, oligomerization, destabilization, and phosphorylation have been reported as candidate biomarkers of storage quality and posttransfusion effects in RBC units . Omics studies have expanded knowledge on the metabolic, biomechanical, and oxidative effects of storage on RBCs and their molecular basis as a function of storage duration, strategy, and probable clinical relevance .…”

A comparison of different methods of red blood cell leukoreduction and additive solutions on the accumulation of neutrophil‐priming activity during storage

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