ABSTRACT

The principle of affinity electrophoresis was successfully used for the quantative determination of molecule interactions. This chapter discusses technical details of agarose crossed-affinity immunoelectrophoresis (CAIE) performed in acidic and neutral pH and details of the CAIE-lectin-inhibition system and provides mathematical equations derived for the determination of dissociation constants based on the experimental data obtained in both systems. The mobility of the serum proteins was retarded when the electrophoresis was carried out in the lectin-containing gel. The reaction of lectin and glycoprotein can be compared to a particular case of enzyme-substrate interaction in which formation of the products is negligible compared to the dissociation of enzyme-substrate complexes. The detailed theory as well as the mathematical equations for calculation of the lectinglycoprotein complexes, dissociation constants by affinity electrophoresis in polyacrylamide gel was developed by V. Horejsi. Similarly, in polyacrylamide gel affinity electrophoresis, Horejsi observed deviation from linearity which was dependent on the glycoprotein concentration.