Characterization of monoclonal antibodies against human thyrotropin and use in an immunoradiometric assay and immunohistochemistry.
Monoclonal antibodies were prepared against human thyrotropin. 13 different antibodies were characterized. Ten antibodies were of the IgG1 subclass. The affinities of the antibodies were in the range 10(9)-10(11) mol-1 X l. Four of them were specific for hTSH and did not react with hLH, hFSH, hCG or alpha hCG. Four reacted with these hormones and recognized the alpha subunit of hCG. One cross-reacted only with hFSH. The remaining four antibodies recognized the holo-hTSH only, and thus were designated as anti-conformational determinants. Monoclonal antibodies reacting with different antigenic determinants on the hTSH molecule defined seven clusters. Two of them were used to develop a simplified two-site sandwich radioimmunoassay in which one monoclonal antibody was immobilized on tubes (anti-beta TSH) and another (anti-alpha) labelled with 125I. This assay was highly specific and demonstrated a sensitivity level of 0.1 microIU/ml. Two monoclonal antibodies were used in immunohistochemistry and their quality and specificity was assessed in the detection of hTSH immunoreactivity in human pituitary biological sections.
D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000918:Antibody Specificity; D006063:Chorionic Gonadotropin / Q000097:blood; D003429:Cross Reactions; D006651:Histocytochemistry; D006801:Humans; D006980:Hyperthyroidism / Q000097:blood; D007037:Hypothyroidism / Q000097:blood; D007457:Iodine Radioisotopes; D011487:Protein Conformation; D011863:Radioimmunoassay / Q000379:methods*; D013972:Thyrotropin / Q000097:blood*
D000911:Antibodies, Monoclonal; D006063:Chorionic Gonadotropin; D007457:Iodine Radioisotopes; D013972:Thyrotropin