Synthesis of Hemoglobin-Based Oxygen Carrier Nanoparticles By Desolvation Precipitation

Hemoglobin (Hb)-based oxygen carriers (HBOCs) present an alternative to red blood cells (RBCs) when blood is not available. However, the most widely used synthesis techniques have fundamental flaws, which may have contributed toward disappointing clinical application. Polymerized Hb contains a heterogeneous distribution of particle size and shape, while Hb encapsulation inside liposomes results in high lipid burden and low Hb content. Meanwhile, there are a variety of other nanoparticle synthetic techniques which, having found success as drug delivery vehicles, may be well suited to function as an HBOC. We synthesized desolvated Hb nanoparticles (Hb-dNPs) with diameters of approximately 250 nm by the controlled precipitation of Hb with ethanol. Oxidized dextran was found to be an effective surface stabilizing agent that maintained particle integrity. In vitro biophysical characterization showed a high-affinity oxygen delivery profile (P₅₀ = 7.72 mm Hg), suggesting a potential for therapeutic use and opening a new avenue for HBOC research.