Controlled Polymerization and Ultrafiltration Increase the Consistency of Polymerized Hemoglobin for Use as an Oxygen Carrier

Polymerized human hemoglobins (PolyhHbs) are a promising class of red blood cell substitute for use in transfusion medicine. Unfortunately, the application of the commonly used glutaraldehyde cross-linking chemistry to synthesize these materials results in a complex mixture of PolyhHb molecules with highly varied batch-to-batch consistency. We implemented a controlled method of gas exchange and reagent addition that results in a homogeneous PolyhHb product. A fully coupled tangential flow filtration system was used to purify and concentrate the synthesized PolyhHb molecules. This improved method of PolyhHb production could be used to more precisely control the size and reduce the polydispersity of PolyhHb molecules, with minimal effects on the resulting oxygen-carrying capability. In addition to these factors, we assessed how the hemoglobin scavenging protein haptoglobin (Hp) would interact with PolyhHb molecules of varying sizes and quarternary states. Our results indicated that T-state PolyhHbs may be more efficiently detoxified by Hp compared with R-state PolyhHb and unmodified Hb.