Abstract
ELISA experiments based on competition between immobilized and soluble antibody for soluble antigen, and on the formation of a ternary complex of immobilized antibody, antigen and soluble antibody were used by Hoylaerts et al. (J. Immunol. Methods 126 (1990) 253–261) for the determination of dissociation constants. The dissociation constant was taken from linearized plots according to a theory that required several approximations. The effect of these approximations on the resulting dissociation constants has been investigated using two computer programs, CBEIA_C and CBEIA_S. Since most approximations that have to be made for linearization can be avoided by non-linear regression, the programs provide a more reliable basis for calculation of the dissociation constant. Experiments measuring formation of a ternary complex were found to be unsuitable for affinity determination.
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