Breast cancer estrogen receptor assays: assessment of a direct hyperbolic fitting to analyse multipoint binding data

Abstract

Analysis of breast cancer estrogen receptor multipoint binding assay is performed by fitting experimental data to a hyperbolic model derived from the law of mass action. The calculations performed on a microcomputer are carried out from the total bound and free ligand concentrations. The parameters estimated by hyperbolic fitting, receptor concentration N and constant of dissociation K, well agree with those obtained by Scatchard's transformation. N and K derived from hyperbolic analysis are much less susceptible to the influence of experimental errors. The method is more reliable at low receptor concentrations. The main advantage of the hyperbolic fitting is to simplify the technical methodology in clinical laboratory practice; there is no need to determine the non-specific bindings experimentally. Calculations can be easily automated on any laboratory microcomputer. Assays of any kind of receptor could be analysed by the hyperbolic fitting when the physical-chemical equilibrium between receptor, nonsaturable component and ligand can be approximated by a two-component model.