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

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.

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 (RCC^WB* ). Two WB units were produced into RCC; one was gamma-irradiated (RCC^γ ) and the other was left untreated as control (RCC^Ø ). In vitro quality parameters were measured during storage. Membrane protein profiles of RCC^Ø , RCC^γ , and RCC^WB* were assessed on selected hemoglobin-depleted membrane fractions using a quantitative proteomics approach based on iTRAQ.

Quantitative proteomic analysis identified 100 proteins at the membrane, with seven unique proteins exhibiting significant changes in RCC^WB* at day 28 of storage. Membrane peroxiredoxin-2, catalase, and proteasome levels demonstrated robust negative correlation with percentage hemolysis.

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.