Integrated manufacturing of REciclable multi-material COmposites for the TRANSport sector
Scalable production and biophysical characterization of an enzyme cocktail derived from human red blood cells
Red blood cells (RBCs) play a critical role in oxygen and carbon dioxide transport, which is facilitated by RBC?encapsulated hemoglobin (Hb) and carbonic anhydrase (CA). In addition, RBCs are constantly exposed to oxidative stress due to the intracellular reactive oxygen species (ROS) generated during Hb auto?oxidation. Antioxidant enzymes within RBCs, such as superoxide dismutase (SOD), catalase (CAT), and peroxiredoxin (Prx), counteract ROS generation to protect the RBC from oxidative stress. Therefore, this study presents a scaled?up method to extract an enzyme cocktail from lysed human RBCs, enriched with the major RBC enzymes with minimal Hb contamination. Using ethanol?chloroform precipitation and multiple biophysical analyses (SDS?PAGE, SEC?HPLC, MALDI?TOF, and LC?MS/MS), the RBC enzymes were successfully separated from Hb in the hemolysate. The purified enzyme cocktail exhibited minimal Hb contamination and retained a significant amount of CA, and antioxidative enzymes like SOD and CAT. Therefore, this scalable RBC enzyme purification method provides an efficient approach for isolating RBC enzymes with broad biomedical relevance.
» Publication Date: 19/09/2025
This project has received funding from the European Union's Horizon 2020 research and innovation programme under grant agreement Nº 768737
