Preparation of Lipid-Free Human Hemoglobin by Dialysis and Ultrafiltration
- 1 June 1987
- journal article
- research article
- Published by Wiley in Biotechnology and Applied Biochemistry
- Vol. 9 (3) , 230-238
- https://doi.org/10.1111/j.1470-8744.1987.tb00474.x
Abstract
Dialysis of human red blood cells using a hypotonic solution and a commercial kidney dialysis unit followed by ultrafiltration through 0.1 .mu.m pore hollow fibers provides an easily managed method for isolation of lipid-free hemoglobin. High pressure liquid chromatography analysis of lipid-free hemoglobin (LFHB) indicates 99-100% protein purity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis demonstrated that LFHB migrates as a single band. The process requires hypoosmotic dialysis of human RBC to a final 119-139 (av 132) mosmol/kg osmotic pressure. Additional reduction in osmotic pressure results in irreversible cell lysis which results in lipid contamination of the hemoglobin. Processing one-half liter of packed red blood cells requires 10 h, resulting in an average of 90% hemoglobin recovery.This publication has 1 reference indexed in Scilit:
- A continuous-flow high-yield process for preparation of lipid-free hemoglobinAnalytical Biochemistry, 1986