A thyroid cancer specific monoclonal antibody which recognizes cryptic epitope(s) of human thyroglobulin

A monoclonal antibody (TCM-9) specific for human thyroid cancer but not for Graves' disease, adenoma or normal thyroid tissue was shown to recognize a 300 K protein but not to bind to native or mature human thyroglobulin (Tg). In this study, we investigated further the antigen recognized by TCM-9. When purified Tg was treated with periodate, dithiothreitol (DTT) and sodium dodecyl sulfate (SDS), immunoblotting of treated Tg with TCM-9 revealed an apparent enhancement of the staining only with DTT-treated Tg. Furthermore, the DTT-treated Tg was shown to bind dose-dependently to plates coated with TCM-9. We performed SDS-PAGE and immunoblot analyses on the immune complex obtained from a homogenate of thyroid cancer tissue which was preincubated with an excess of anti-Tg monoclonal antibody. When the protein was autoradiographed with [125I]TCM-9, a definite band was observed. The results indicate that TCM-9 is likely to be directed against a masked epitope of thyroglobulin which can be exposed after treatment with a reducing agent.