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

C. L. Sheffield Veterinary Toxicology and Entomology Research Laboratory Agricultural Research Service, United States Department of Agriculture College Station, Texas, 77841

G. E. Spates Veterinary Toxicology and Entomology Research Laboratory Agricultural Research Service, United States Department of Agriculture College Station, Texas, 77841

J. R. Deloach Veterinary Toxicology and Entomology Research Laboratory Agricultural Research Service, United States Department of Agriculture College Station, Texas, 77841

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.

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

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

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