Purification of hemoglobin from red blood cells using tangential flow filtration and immobilized metal ion affinity chromatography
2011
Elmer, Jacob | Harris, David | Palmer, Andre F.
Two methods for purifying hemoglobin (Hb) from red blood cells (RBCs) are compared. In the first method, red blood cell lysate is clarified with a 50nm tangential flow filter and hemoglobin is purified using immobilized metal ion affinity chromatography (IMAC). In the second method, RBC lysate is processed with 50nm, 500kDa, and 50–100kDa tangential flow filters, then hemoglobin is purified with IMAC. Our results show that the hemoglobins from both processes produce identical Hb products that are ultrapure and retain their biophysical properties (except for chicken hemoglobin, which shows erratic oxygen binding behavior after purification). Therefore, the most efficient method for Hb purification appears to be clarification with a 50nm tangential flow filter, followed by purification with IMAC, and sample concentration/polishing on a 10–50kDa tangential flow filter.
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