Identification of potential protein quality markers in pathogen inactivated and gamma-irradiated red cell concentrates.

PURPOSE: Post-collection manipulations (PCMs) aim to increase blood product safety. However, PCMs improve safety at a cost to quality, causing elevated hemolysis. As hemolysis is linked to red blood cell membrane integrity, a quantitative proteomics approach was employed to assess membrane proteome alterations induced by PCMs. EXPERIMENTAL DESIGN: Three ABO-matched whole blood (WB) units were pooled-and-split into three identical units. One WB unit was treated with riboflavin/ultraviolet illumination prior to red cell concentrate (RCC) production (RCCWB* ). Two WB units were produced into RCC; one was gamma-irradiated (RCCγ ) and the other was left untreated as control (RCCØ ). In vitro quality parameters were measured during storage. Membrane protein profiles of RCCØ , RCCγ , and RCCWB* were assessed on selected hemoglobin-depleted membrane fractions using a quantitative proteomics approach based on iTRAQ. RESULTS: Quantitative proteomic analysis identified 100 proteins at the membrane, with seven unique proteins exhibiting significant changes in RCCWB* at day 28 of storage. Membrane peroxiredoxin-2, catalase, and proteasome levels demonstrated robust negative correlation with percentage hemolysis. CONCLUSION: Overall, the in vitro parameters and alterations of membrane protein profiles indicated that pathogen inactivation treatment impacts RCC quality more severely than gamma-irradiation and that it may induce damage through a predominately oxidative mechanism.