Electrophoretic Studies of Thyroxine and Triiodothyronine Binding by the Sera of Guinea Pigs Immunized Against Thyroglobulin

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Abstract
The distribution of radiothyroxine and radiotriiodothyronine in the electrophoretic pattern of normal and thyroglobulin- immune guinea pig sera was studied by both agar and paper electrophoresis. In normal or only adjuvant-treated animals, both thyroxine and triiodothyronine migrated to the albumin region. With thyroglobulin-immune sera, however, an additional peak of radioactivity was obtained in the beta-gamma globulin region as well as the one in the albumin region. The area covered by the peak of radioactivity in the beta-gamma globulin region differed in individual guinea pigs, presumably in relation to the level of circulating antibody, and could be inhibited by pretreating the immune serum with stable thyroxine. A precipitation reaction was obtained by layering a thyroglobulin solution onto immune serum but no precipitation reaction could be elicited with thyroxine or triiodothyronine. No radioactivity peak in the beta-gamma globulin region could be obtained with I131- labeled diiodotyrosine, radio-Rose Bengal or Insulin-I131 mixed with thyroglobulin-immune sera; nor was such a radioactivity peak obtained when radioactive thyroxine was mixed with an insulin-immune serum. The significance of the binding of thyroid hormones by thyroglobulin-immune sera has been discussed.