Immobilized metal ion affinity gel electrophoresis: Quantification of protein affinity to transition metal chelates

This paper describes some recent advances in the methodology of immobilized metal ion affinity gel electrophoresis. Four different ways to incorporate metal chelate ligands in agarose and polyacrylamide-based electrophoresis gels are evaluated, a new polymerizable metal chelating ligand, allyl-2-hydroxy-3-(N,N-dicarboxymethyl)amino-propyl ether, is introduced, and the determination of affinity constants described. The affinities of model proteins (ribonucleases A and B and cytochromes c from different species) for the transition metal chelate iminodiacetic acid-Cu(II) were studied. The results were found to be in agreement with literature data on immobilized metal ion affinity chromatography, and the polymer nature and the different chemistries used influenced the affinity only quantitatively, keeping the basic mechanisms of interaction unchanged.