Effect of ligand‐affinity differences of human hemoglobin variants on electrophoretic behavior and their isolation and functional characterization

Abstract

A natural sulfated polysaccharide (agaropectin), contained in crude agar, can be used as a medium for electrophoretic separation of hemoglobin mutants, constituting a particular class of protein-ligand interactions. Mutations which either modify the electrostatic charge at the surface of the hemoglobin molecule or not, have been studied according to their putative interaction with the medium. Using conformational specificities of the hemoglobin molecule, we have also demonstrated that isoelectric focusing on a polyacrylamide gel in the absence of heme ligands represents a useful, convenient and rapid procedure for isolating silent Hb variants in their native form, provided that they exhibit an abnormal Bohr effect.

Affinity in Electrophoresis

“…In its lifetime a substantial number of papers and reviews dealing with affinity interactions in electrophoretic techniques have been published in Electrophoresis (Table 1). More than 125 papers 18, 38, 46, 47, 49–53, 69–71, 93, 101, 110, 118, 129–238 with the word affinity in the title and dealing with some aspect of affinity electrophoresis have been published in the journal since 1989, the year it started being indexed in PubMed. Prior to that, i.e.…”