Solid-phase radioimmunoassay of thyroxine in untreated serum

We describe a simple, convenient solid-phase radioimmunoassay of total thyroxine in unextracted serum. Serum samples are added directly to the assay incubation mixture, interference in the antigen/antibody reaction by the thyroxine-binding serum proteins being almost completely eliminated by the addition of 8-anilino 1-naphthalene sulfonic acid and incubation at pH 10.5. Residual interference is compensated for by including thyroxine-free serum in the standards. Use of thyroxine antibodies that are coupled to a solid support permits separation of free and antibody-bound hormone by a single washing step, followed by centrifugation. The method is specific, accurate, and reasonably precise. The results obtained compare well with those for radioimmunoassay of thyroxine in serum freed of protein by gel filtration, and with results of a competitive protein-binding method. The technical simplicity of the procedure should readily permit automation. These features suggest that the technique should be well suited for routine clinical laboratory use.