抗人甲状腺球蛋白单克隆抗体作为甲状腺球蛋白结构的探针

Monoclonal antibodies to human thyroglobulin as probes for thyroglobulin structure.

作者信息

De Baets M H, Theunissen R, Kok K, de Vijlder J J, van Breda Vriesman P J

出版信息

Endocrinology. 1987 Mar;120(3):1104-11. doi: 10.1210/endo-120-3-1104.

Abstract

A panel of monoclonal antibodies (mAbs) against human thyroglobulin (hTg) was obtained by somatic fusion of the nonsecreting myeloma cell line P3X66 Ag8/0 and spleen cells of Balb/c mice immunized with purified hTg. Antibody secreting clones were selected by solid phase enzyme immunoassay and analyzed for cross-reaction with Tg from several animal species. Twelve out of 15 mAbs cross-reacted with both rat and mouse Tg and 11 Mabs cross-reacted with bovine Tg. The cross-reaction with mouse Tg paralleled that of rat Tg, whereas discrete differences between the cross-reactivity patterns with bovine Tg were observed. Two clones secreted mAbs specific for hTg. We further characterized the mAbs and found that three mAbs recognized T4-containing determinants and one mAb reacted with both T4 and T3-containing determinants on the Tg molecule. The binding of the mAbs to hormonogenic determinants depended upon the thyroid hormone content of the molecule and the integrity of the three dimensional structure of Tg. One other mAb reacted with four peptides of CNBr-cleaved hTg, indicating the recognition of a repetitive determinant in the hTg molecule distinct from the hormonogenic regions.

摘要

通过非分泌性骨髓瘤细胞系P3X66 Ag8/0与用纯化的人甲状腺球蛋白（hTg）免疫的Balb/c小鼠的脾细胞进行体细胞融合，获得了一组抗人甲状腺球蛋白（hTg）的单克隆抗体（mAb）。通过固相酶免疫测定法筛选出分泌抗体的克隆，并分析其与几种动物物种的甲状腺球蛋白（Tg）的交叉反应。15种单克隆抗体中有12种与大鼠和小鼠的Tg发生交叉反应，11种单克隆抗体与牛的Tg发生交叉反应。与小鼠Tg的交叉反应与大鼠Tg的交叉反应相似，而与牛Tg的交叉反应模式之间存在明显差异。两个克隆分泌对hTg特异的单克隆抗体。我们进一步对这些单克隆抗体进行了表征，发现三种单克隆抗体识别含T4的决定簇，一种单克隆抗体与Tg分子上含T4和T3的决定簇都发生反应。单克隆抗体与激素生成决定簇的结合取决于分子的甲状腺激素含量和Tg三维结构的完整性。另一种单克隆抗体与CNBr裂解的hTg的四种肽段发生反应，表明识别出hTg分子中不同于激素生成区域的重复决定簇。