Analysis and Application of an Equilibrium Model forin vitroBioassay Systems with Three Components: Receptor, Hormone and Hormone-Binding-Protein

A simple theoretical framework is presented for bioassay studies using three componentin vitrosystems. An equilibrium model is used to derive equations useful for predicting changes in biological response after addition of hormone-binding-protein or as a consequence of increased hormone affinity. Sets of possible solutions for receptor occupancy and binding protein occupancy are found for typical values of receptor and binding protein affinity constants. Unique equilibrium solutions are dictated by the initial condition of total hormone concentration. According to the occupancy theory of drug action, increasing the affinity of a hormone for its receptor will result in a proportional increase in biological potency. However, the three component model predicts that the magnitude of increase in biological potency will be a small fraction of the proportional increase in affinity. With typical initial conditions a two-fold increase in hormone affinity for its receptor is predicted to result in only a 33% increase in biological response. Under the same conditions an 11-fold increase in hormone affinity for receptor would be needed to produce a two-fold increase in biological potency. Some currently used bioassay systems may be unrecognized three component systems and gross errors in biopotency estimates will result if the effect of binding protein is not calculated. An algorithm derived from the three component model is used to predict changes in biological response after addition of binding protein toin vitrosystems. The algorithm is tested by application to a published data set from an experimental study in anin vitrosystem (Limet al., 1990,Endocrinology127,1287–1291). Predicted changes show good agreement (within 8%) with experimental observations.