One-Step, Labeled-Antibody Assay for Measuring Free Thyroxine. 1. Assay Development and Validation

Abstract

We describe a one-step, labeled-antibody radioassay for measuring free thyroxin (FT4) in serum or plasma, based on a novel principle. FT4 in the sample competes with a gross molar excess (over antibody) of a cross-reactant (L-triiodothyronine, T3), chemically coupled to magnetizable polymer particles, for binding to avid 125l-labeled monoclonal anti-thyroxin antibodies. As in conventional immunoassays, 125I counts bound to the solid phase (T3-magnetizable particles) are inversely proportional to sample FT4 concentration. We demonstrate here the development and technical validity of this new method.