Radioimmunoassays of 3,5,3'-triiodo-L-thyronine with and without a prior extraction step

Abstract

Two radioimmunoassays for triiodothyronine (T3) are described, one of which includes an extraction step, while the other does not. To raise antibodies, two carrier proteins and different coupling agents were used, namely haemocyanin and diazotized benzidine or human serum albumin and carbodiimide. In the case of T3 coupled to haemocyanin by diazotized benzidine, evidence of covalent binding of the hapten to the protein was obtained. In the case of T3 coupled to human serum albumin, little evidence of covalent linkage was available. Nevertheless immunization was successful in both cases. The radioimmunoassay in unextracted serum was highly reproducible and precise (intra-assay variability 5.2% inter-assay variability 8.1%). Normal values were determined which clearly indicate a fall in the serum T3 concentration with increasing age. In men the fall occurs in the fifth decade. In women the T3 starts to fall only after 70 years of age. In 31 cases of hyperthyroidism the serum T3 concentration ranged from 2.26 to 10.4 ng T3/ml. In 10 cases of hypothyroidism the values ranged from 0 to 0.8 ng T3/ml. The radioimmunoassay using an extraction procedure was less extensively used since it was found to be less reproducible (intra-assay variability 7.5%, inter-assay 12.25%). The normal values were determined with a mixed population aged 20-50. The mean +/- 2 SD was 0.9 /- .36 ng T3/ml (n = 52). In 17 cases of hypothyroidism the values ranged from 0 to 0.6 ng T3/ml and in 22 cases of hyperthyroidism from 2 to 14.4 ng T3/ml.