Development of Zinc Chelating Resin Polymer Beads for the Removal of Cell-Free Hemoglobin

Abstract

Objective: Red blood cell (RBC) hemolysis is one of the most common storage lesions in packed RBCs (pRBC). Older units of pRBCs, especially those >21 days old, have increasing levels of hemolysis leading to increased oxidative stress and premature platelet activation. This effect can mostly be attributed to the increase of cell-free hemoglobin (Hb). Therefore, removal of cell-free Hb from pRBCs prior to transfusion could mitigate these deleterious effects. We propose a new method for the removal of Hb from pRBCs using zinc beads.Approach and Results: Prepared Hb solutions and pRBCs were treated with zinc beads using two different protocols. UV-Vis spectrophotometry was used to determine Hb concentrations, before and after treatment. Experiments were run in triplicate and paired t-tests were used to determine significant differences between groups. Zinc beads removed on average 94% of cell-free Hb within 15 minutes and 78% Hb from pRBCs (p<0.0001), demonstrating a maximum binding capacity ~66.2±0.7 mg Hb/mL beads. No differences in RBC morphology or deformability were observed after treatment. Conclusion:This study demonstrates the feasibility of using zinc beads for the rapid and targeted removal of Hb from pRBC units. Further investigation is needed to scale this method for large volume removal.