Development of Detection System for Thyroid Carcinoma Specific Antigen and its Function Analysis

本研究旨在鉴别甲状腺乳头状癌。我们首先开发了可以特异性识别甲状腺肿瘤相关抗原的单克隆抗体，这使我们能够通过蛋白质印迹和ELISA研究该抗原在各种组织中的表达。我们研制了两种类型的单克隆抗体，如甲状腺癌抗体（CAb）和甲状腺球蛋白（Tg）抗体（TgAb）。使用TgAb，基于先前观察到甲状腺癌相关Tg和正常Tg中的糖链与凝集素的反应性存在差异，建立了区分甲状腺癌组织和正常甲状腺组织的ELISA系统。在使用CAb的Western印迹中，在所有乳头状癌相关组织标本中观察到约40 kDa的蛋白。相反，在其他甲状腺组织中从未检测到40 kDa条带。随后，使用LCA凝集素通过ELISA研究各种甲状腺疾病中未结合血清Tg的比率{Tg-unbound（%）}。甲状腺乳头状癌的TG-未结合（%）显著高于良性甲状腺肿和正常对照组。这些结果清楚地表明，单克隆抗体为基础的蛋白质印迹和凝集素为基础的ELISA将显着有助于甲状腺乳头状癌患者的诊断。对40 kDa蛋白质具有特异性的单克隆抗体正在开发中。

The aim of this study was to differentiate thyroid papillary carcinoma from the other. We first developed monoclonal antibodies that could specifically recognize a thyroid tumor-associated antigen, which enabled us to investigate the expression of this antigen in various tissues by western blotting and ELISA. We developed two types of monoclonal antibodies such as thyroid carcinoma antibodies (CAb) and thyroglobulin (Tg) antibodies (TgAb). Using the TgAb, an ELISA system was established to differentiate thyroid cancer tissues from normal thyroid tissue, based on the previous observation that there are differences in the reactivity with lectins between the sugar chains in thyroid cancer associated Tg and normal Tg. In western blotting using CAb, a protein of about 40kDa was observed in all the papillary carcinoma associated tissue specimens. In contrast, the 40kDa band was never detected in any of the other thyroid tissues. Subsequently, the ratios of unbound serum Tg {Tg-unbound (%)} in various thyroid diseases were investigated by ELISA using LCA lectin. The Tg-unbound (%) in papillary carcinoma was significantly higher than that observed in benign goiter and in normal controls. These results clearly indicate that the monoclonal antibody-based western blotting and lectin-based ELISA would significantly contribute to diagnose patients with thyroid papillary carcinoma. Monoclonal antibodies specific for the 40 kDa protein is under development.