Interference of anti-T4 autoantibodies in the single step free T4 radioimmunoassays

Abstract

The thyroid hormone profile of a hypothyroid patient, with moderate sized diffuse goiter and history of exogenous thyroid hormone therapy, was found to be highly ambiguous. The total as well as free T4 levels were consistently high (out of range) with suppressed T3 (total and free) and elevated thyrotropin levels. We incubated the serum with the radio-iodine labelled T4 and observed that the patient's serum bound about 50.5% of the total radioactivity resulting in 85.3% interference in the radioimmunoassay. On resolving the serum proteins by cellulose acetate electrophoresis, the radioactivity was detected in the gamma globulin band. The protein profile showed mildly raised levels of IgG. Therefore it was established that the patient's serum carried T4 reactive antibodies (IgG) which were interfering in the immunoassays. Elevated anti-thyroid microsomal antibodies were also present in the patient's serum. The anti-T4 antibodies were highly specific for T4 and did not show any cross reactivity with the T3 or its analogues used in the estimation of free T3. We stripped the serum by precipitating the immunoglobulins and were successful in estimating the levels of free T4 in the stripped serum by single step radioimmunoassy.