Automatic treatment of radioimmunoassay data: an experimental validation of the results

A computational method for the automatic treatment of radioimmunoassay data has been developed and a computer program has been written in accordance. The main features of the approach used are the following: (a) a constant ratio between the bound fraction and its variance is assumed and estimated in each assay; (b) the points of the dose-response curve are fitted using the three-parameter function y' equals b1/(1 plus b2x(-b)3) where y' equals bound - nonspecific counts and x is the amount of hormone; the fitting is performed using the nonlinear, least-squares technique; (c) the values of the unknown samples are evaluated from the fitted standard curve; their confidence limits are computed taking into account both the variance of the bound replicates and the variance of the parameters of the dose-response curve. Experimental data that support the validity of the assumption on the variance of bound measurement and the suitability of the chosen function to fit the points of the standard curve are presented. A comparison between the confidence limits of the unknowns experimentally obtained and those computed by the program is reported and discussed.