Related Experiment Video

Updated: Jun 5, 2026

Staphylococcus aureus Growth using Human Hemoglobin as an Iron Source
06:37

Staphylococcus aureus Growth using Human Hemoglobin as an Iron Source

Published on: February 7, 2013

Purification of hemoglobin from red blood cells using tangential flow filtration and immobilized metal ion affinity

Jacob Elmer1, David Harris, Andre F Palmer

  • 1William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 West 19th Avenue, Columbus, OH 43210, USA.

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
|January 4, 2011
PubMed

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 50 nm tangential flow filter and hemoglobin is purified using immobilized metal ion affinity chromatography (IMAC). In the second method, RBC lysate is processed with 50 nm, 500 kDa, and 50-100 kDa 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 50 nm tangential flow filter, followed by purification with IMAC, and sample concentration/polishing on a 10-50 kDa tangential flow filter.

More Related Videos

Purification of the M. magneticum Strain AMB-1 Magnetosome Associated Protein MamAΔ4111:07

Purification of the M. magneticum Strain AMB-1 Magnetosome Associated Protein MamAΔ41

Published on: March 25, 2010

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain14:25

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain

Published on: December 12, 2017

Related Concept Videos

Extracorporeal Removal of Drugs: Hemoperfusion and Hemofiltration01:25

Extracorporeal Removal of Drugs: Hemoperfusion and Hemofiltration

Hemoperfusion and hemofiltration are critical techniques in medical treatments to eliminate accumulated drugs, metabolites, and electrolytes from the bloodstream. These methods are particularly vital in cases of accidental poisoning and drug overdose.Hemoperfusion involves passing blood through an adsorbent material to remove unwanted substances. The main adsorbents used in hemoperfusion include activated charcoal and Amberlite resins. Activated charcoal can adsorb both polar and nonpolar...
Affinity Chromatography01:03

Affinity Chromatography

Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
Types Of Column Chromatography01:29

Types Of Column Chromatography

The stability and compatibility of column material with samples are crucial for efficient purification in chromatographic techniques. Various operating parameters such as pH, temperature, or solvent affect the packing of the column material, thereby determining the purification efficiency. The choice of column material also plays an essential role in deciding the operating parameters and can be modified based on the proteins that need to be purified.
Gel Filtration Chromatography
When the...
Downstream Processing01:29

Downstream Processing

Downstream processing begins once fermentation is complete and involves a series of steps to recover and purify products such as acids, vitamins, antibiotics, or proteins.Cell HarvestingFor example, for intracellular protein-based products, the first step is harvesting the cells. This is typically achieved using centrifugation or filtration to separate the cells from the liquid phase.Cell Disruption for Intracellular ProductsIf the target product is intracellular, the harvested cells must be...