Specificity of monoclonal antibodies against human thyroglobulin; comparison with autoimmune antibodies

• Published in: The EMBO journal 1983-01, Vol.2 (10), p.1821-1826
• Main Authors: Ruf, J., Carayon, P., Sarles‐Philip, N., Kourilsky, F., Lissitzky, S.
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, immunoglobulins; Antibodies, Monoclonal; Antigen-Antibody Complex; Autoantibodies; Biological and medical sciences; Dogs; Epitopes - analysis; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Haplorhini; Humans; Hybridomas; Mice; Mice, Inbred BALB C; Molecular immunology; monoclonal antibodies; Species Specificity; Swine; thyroglobulin; Thyroglobulin - analysis; Thyroglobulin - immunology
• ISSN: 0261-4189; 1460-2075
• DOI: 10.1002/j.1460-2075.1983.tb01664.x

Ten monoclonal antibodies (mAb) directed against human thyroglobulin (hTgb) were produced, purified and characterized. The mAb avidity for hTgb ranged from 10(‐10) to 10(‐6) M. The species specificity of the mAb was as follows: eight mAb reacted with monkey Tgb, three with dog Tgb and one with pig Tgb; none with bovine and ovine Tgb. The binding of mAb to hTgb was not significantly inhibited in the presence of Tgb carbohydrate moieties, tyrosine, iodotyrosines and iodothyronines. The topology of the antigenic determinants recognized by the 10 mAb on hTgb was explored by inhibition of Tgb binding of radiolabeled mAb by the other antibodies. Six distinct clusters of reactivity were described. Localization of the antigenic determinants recognized by mAb on hTgb was attempted using tryptic fragments of hTgb to inhibit the binding of mAb to hTgb. The inhibitory effect of hydrolysis products was different for each mAb but exhibited partial analogies between mAb of the same cluster of reactivity. Anti‐hTgb autoimmune antibodies (aAb) purified from sera of Graves patients cross‐reacted essentially with mAb of one out of the six clusters. These results demonstrate that the large number of antigenic determinants presented by the hTgb are not disseminated on the molecule but are clustered in antigenic regions. Furthermore, from the six antigenic regions evidenced in this paper, only one is involved in autoimmune antibody production in Grave's disease.