Radial partition immunoassay applied to automated quantification of human choriogonadotropin with use of two monoclonal antibodies.

We describe a novel application of radial partition immunoassay to quantification of human choriogonadotropin (hCG). In this "sandwich"-type assay, two monoclonal antibodies, specific for different sites on the intact molecule are used. The solid phase consists of tabs of glass-fiber filter paper containing a pre-immobilized antibody specific for the alpha subunit of hCG. The patient's sample is first applied directly to the central "reaction zone" of the tab, allowing hCG to bind to the solid-phase antibody. Then a buffered solution containing enzyme-labeled Fab' fragments of a monoclonal antibody specific for the beta subunit of hCG is applied, initiating "sandwich" formation. Finally, a wash buffer containing a fluorogenic substrate is applied, eluting unbound conjugate to the tab periphery. Bound enzyme conjugate is quantified by measuring the rate of increase in fluorescence. Rates are converted to clinical units by comparison with a stored calibration curve. Elapsed time from sample application to results is less than 8 min. Specific performance characteristics of this assay are reported.