Progress in affinophoresis

Abstract

The use of polyionic polymers as mobile affinity matrices in electrophoresis has led to the development of a specific separation method for biological substances, affinophoresis. The conjugate of a polyionic polymer and an affinity ligand is called an affinophore. Electrophoresis of proteins in the presence of an affinophore results in a change in the mobility of a specific protein due to the difference between the mobility of the protein and that of the protein-affinophore complex. Polylysine is useful as a base polymer of affinophores and has been used successfully as an anionic matrix after succinylation. Affinophoresis of proteases, lectins and antibodies has been carried out in agarose gel and the mobility of the protein having affinity to each ligand was specifically changed. Two-dimensional affinophoresis, in which an affinophore was included only in the second-dimensional electrophoresis, was effective for the separation of the components of a complex mixture of proteins even if the change of mobility was not large. Red blood cells were successively treated with homologous antiserum, biotinylated second antibody, avidin and biotinylated succinylpolylysine as an affinophore. Specific acceleration of the homologous cells to the antiserum was observed even when the affinophoresis was applied to mixed red blood cells from different species.