PMID: 3766962Aug 15, 1986Paper

A continuous-flow high-yield process for preparation of lipid-free hemoglobin

Analytical Biochemistry
J R DeLoachG E Spates

Abstract

Hypotonic hollow-fiber dialysis of bovine red blood cells followed by ultrafiltration through 0.1-micron 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-polyacrylamide 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.

References

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Citations

Nov 30, 1992·Clinica Chimica Acta; International Journal of Clinical Chemistry·P T BiesselsJ C Bakker
Jan 1, 1994·Artificial Cells, Blood Substitutes, and Immobilization Biotechnology·P Kan, C J Lee
Nov 17, 2009·Methods in Enzymology·Hiromi SakaiEishun Tsuchida
Jan 1, 1994·Artificial Cells, Blood Substitutes, and Immobilization Biotechnology·H SakaiE Tsuchida
Jan 1, 1988·Biomaterials, Artificial Cells, and Artificial Organs·C L SheffieldJ R DeLoach
Jan 1, 1992·Biomaterials, Artificial Cells, and Immobilization Biotechnology : Official Journal of the International Society for Artificial Cells and Immobilization Biotechnology·C J LeeW K Chen
Jan 1, 1988·Biomaterials, Artificial Cells, and Artificial Organs·W M VogelC R Valeri
Jul 28, 2004·Artificial Cells, Blood Substitutes, and Immobilization Biotechnology·Xiuling LuZhiguo Su

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