Studies on the Biospecific Molecular Interactions by Means of Affinity Electrophoresis

The principle of the affinity electrophoresis was summarized. Three affinity equations, the original, the inhibition, and the general affinity equations, were drived from the affinity theory. According to those equations, dissociation constants of various enzymes for high and low molecular substrates, lectins for oligo- and polysaccharides, anti-dextran myeloma proteins for dextran and isomaltose oligosaccharides, and anti-DNP-antibodies for DNP-polyacrylamide copolymer were determined. For highly heterogeneous protein mixtures, the two-dimensional affinity electrophoresis showed an excellent resolution. Thus, rabbit anti-DNP-antibodies were separated into over 200 spots on one affinity gel. Dissociation constants for every IgG spot were directly calculated from the affinity pattern. Effects of temperature and pH on the affinity electrophoresis were discussed.