HOMOGENEOUS 2-SITE IMMUNOMETRIC ASSAY KINETICS AS A THEORETICAL TOOL FOR DATA-ANALYSIS

The easily accessible kinetics of a new homogeneous two-site fluoromet ric immunoassay for prolactin was studied, in order to determine its u sefulness for assay data reduction and optimization. The combined use of a simple descriptive model fitted to experimental data and a mechan istic model to simulate the kinetics revealed that (i) the kinetics cu rve presented an early inflexion point. Its time of occurrence was con stant as long as the antigen concentration was below the smallest anti body concentration and decreased to zero for higher concentrations. It may therefore be used as an indicator of hooked samples. (ii) The kin etics steepest slope was correlated with antigen concentration, Its us e as a dose-response curve variable would allow higher concentrations to be assayed than with the classical end-point dose-response curve. T he results suggest that control and exploitation of kinetic parameters could help to improve the rapidity, analytical range, and reliability of homogeneous two-site immunometric assays. (C) 1997 Academic Press.