Abstract
Hypotonic hollow-fiber dialysis of bovine red blood cells followed by ultrafiltration through 0.1-μm pore hollow fibers provides a simple method for isolation of lipid-free hemoglobin. Hemoglobin (Hb) isolated by comparative techniques were all contaminated with membrane stroma. HPLC analysis of Hb revealed a protein peak of 99.6% purity and sodium dodecylsulfate-poly-acrylamide gel electrophoresis analysis revealed a single band. The process requires hypoosmotic dialysis of bovine RBC to a final 160–180 mosmol/kg osmotic pressure. Additional reduction in osmotic pressure causes irreversible cell lysis which leads to lipid contamination of the Hb. Processing of 1 2 liter of packed red blood cells requires 4–5 h, resulting in an average of 90% hemoglobin recovery.
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