Production and in Vitro Utilization of Monoclonal Antibodies to Human Thyroglobulin

Abstract

Human thyroglobulin (Tg) was used as an antigen in the development of antibodies by the hybridoma technique. From four antibodies that bound more than 40% labeled Tg, two were characterized (182/E4 and 211/A5). They were both of the immunoglobulin G 2ab subclass, and provided an affinity constants (K_a) of 1.2 × 10¹⁰ and 7.7 × 10⁹ mol^-1, respectively. The specificity of these antibodies was demonstrated by the absence of cross-reaction by monoiodothyronine, diiodothyronine, T₃, T₄, and sialic acid. A RIA was developed with 182/E4 or 211/A5, and the least detectable dose, based on the standard curve, was 10 ng/ml. The immunoreactivities of 182/E4 and 211/A5 to four Tg preparations different in iodine content appeared to be identical. Histochemical staining was used on normal and neoplastic tissues with both antibodies. Positive reactions were obtained in both cells and colloid, with heterogeneous staining from one follicle to another. Papillary carcinoma showed numerous positive cells, in contrast with Hürtle cell tumors which displayed very few positive cells. Anaplasic giant and spindle cells were negative. Monoclonal antibodies to human Tg are useful for in vitro detection of Tg. (J Clin Endocrinol Metab56: 530, 1983)