抗人甲状腺球蛋白单克隆抗体的制备

PREPARATION OF MONOCLONAL ANTIBODIES AGAINST HUMANTHYROGLOBULIN

目的：制备抗人甲状腺球蛋白（Ｔｇ）单克隆抗体，为建立高灵敏度的Ｔｇ检测方法做准备。方法：从人的甲状腺组织中提取Ｔｇ做抗原，经腹腔免疫ＢＡＬＢ／ｃ小鼠，取其脾细胞与小鼠骨髓瘤细胞系ＳＰ２／ｏ融合，筛选及克隆化。结果：获得２株分泌抗人Ｔｇ单克隆抗体的杂交瘤细胞株。鼠腹水抗体效价达１．０２×１０６。经免疫球蛋白类别及亚类的鉴定，２株单克隆抗体均为ＩｇＧ１亚类，轻链为Ｋ型。结论：我们制备的抗Ｔｇ单克隆抗体具有良好的免疫反应特性。

Objective:The study described the preparation of monoclonal antibody (McAb) to thyroid globulin(Tg) which can be used for establishing highly sensitive assay for Tg Methods:BALB/c mice were immunized by intraperitioneal injection of Tg from human thyroid glandSpleen cells from the immunized mice were fused with SP2/0 myeloma cell lineThe hybridoma cell line were screened and cloned Results: Two hybridoma cell lines produced McAb to Tg The titer in ascites is 102×106McAb secreted by two hybridoma cell lines were IgG1 subclass and light chain was K type by determining IgG isotype and subclassConclusion:McAb to Tg is good in immune reactivity and is suitable for establishing enzymelined immunosorbent assay (ELISA)