Conclusions
This study reports a new method for hemoglobin deoxygenation and
compares the reaction timescales and magnetic velocities between three
RBC treatment methods on healthy and sickled blood. Addition of enzymes
used in the ETC makes possible the reversible deoxygenation of RBCs at
ambient conditions with high purity of deoxyHb. CTV results confirm that
healthy RBCs have similar paramagnetic behavior compared to accepted RBC
treatments. Additionally, this enzymatic treatment works well with SCD
blood, collected as apheresis waste, which has a similar amount of
intracellular Hb as healthy donors. Intracellular iron calculations from
measurements of RBCs in medium of different viscosities further confirm
the CTV system accuracy. Further, AS-3 storage solution can be buffered
to the optimum pH for Oxyrase activity, without the increased viscosity
when using lactate. Oxygen-dissociation curves show that RBC
interactions with these enzymes do not significantly affect the oxygen
binding properties, suggesting this may be a viable step in clinical
blood processing in the future. In future works the authors wish to
observe the effects of maintaining low DO over the course of a donated
blood unit’s storage life (6 weeks) and fractionate healthy and SCD
blood.