Monoclonal antithyroglobulin antibodies derived from immunizations of mice with human eye muscle and thyroid membranes

A mouse hybridoma clone secreting an immunoglobulin M monoclonal antibody (K-5-4), which reacted with human thyroglobulin (Tg), was obtained from spleen cells of mice immunized with crude membranes of human eye muscle tissues (Em). Its binding to Tg could be inhibited by another monoclonal anti-Tg (Fl-11-1) derived from spleen cells of mice immunized with human thyroid cell membranes, but K-5-4 did not inhibit the binding of Fl-11-1 to Tg. This finding suggests that K-5-4 may react with a site on the Tg molecule which is susceptible to conformational changes, such as that induced by binding of another anti-Tg antibody at another site on Tg. K-5- 4 reacted with human, mouse, rat, guinea pig, bovine, and porcine Tg. Bindingand immunohistological staining experiments failed to detect binding of K-5-4 toEm tissue. The very low frequency of one Tg-reacting hybridoma from 6 X 108 spleen cells fused after Em immunization contrasts with the relative ease with which monoclonal anti-Tgs were generated from spleen c...