Hypoosmotic Dialysis and Ultrafiltration Technique for Preparation of Mammalian Hemoglobin: a Comparison of Three Species

Abstract

Dialysis of human, bovine, and ovine red blood cells using a hypotonic solution and a commercial kidney dialysis unit, followed by ultrafiltration through 0.1μm pore hollow-fibers provides an easily managed method for isolation of lipid-free hemoglobin (LFHB). High performance liquid chromatography (HPLC) analysis of hemoglobin indicated 99% protein purity. SDS polyacrylamide gel electrophoresis analysis demonstrated that LFHB migrates as a single band. Processing one-half liter of packed red blood cells requires approximately 10 hours and resulting in an average of 93% hemoglobin recovery.