Immunologic and Biologic Properties of Thyrotropin Antiserums1

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Abstract
The antigenicity of several bovine thyrotropin preparations (Armour, Pierce, Morris) was demonstrated in rabbits and guinea pigs. Antibody titers determined by the tanned red cell hemagglutination assay (TRC-H) averaged 1:650,000 in guinea pigs and 1:80,000 in rabbits. Column electrophoresis and chromatography of rabbit antiserum showed that the precipitating antibodies migrated with the gamma globulins. Studies on the immunologic specificity of thyrotropin antiserum by the TRC-H and tanned red cell hemagglutination-inhibition (TRC-I) assays, gel diffusion techniques and precipitation and complement fixation reactions suggest some cross-reactivity with other pituitary hormones but indicate that the major antibody constituent is directed against thyrotropin. Inhibition of the biologic action of thyrotropin was shown in assays of thyrotropin in rats (Querido assay) and in mice (McKenzie assay). Application of the hemagglutination- inhibition assay to quantitation in buffer or isotonic saline solutions permitted detection of thyrotropin in amounts of 0.02–0.04 mU. Satisfactory quantitation by this technique was applied to purified bovine thyrotropin (Pierce, Morris) as well as to a preparation obtained by percolation from human thyrotoxic serum (Bates). Inhibitory factors present in human serum and urine interfere with the detection of thyrotropin in biologic media by this assay.