人甲状腺过氧化物酶单克隆抗体的制备与鉴定

DEVELOPMENT AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES TO HUMAN THYROID PEROXIDASE

以亲和纯化的人甲状腺过氧化物酶作免疫原,用经典的杂交瘤技术制备出29株抗人甲状腺过氧化物酶单克隆抗体（TPOmAb）,探讨了甲状腺微粒体抗体（TMAb）与TPObmAb之间的关系。发现TMAb对TPObmAb与^（125）I-hTPO的结合抑制性呈显著的量-效关系（r=0.982,P<0.001）;经间接免疫荧光技术证实原代培养的人甲状腺细胞膜表面存在hTPO,而FRTL-5细胞膜表面不存在hTPO,说明所制备的TPObmAb为hT-PO特异性单克隆抗体。

Purified hTPO was used to prepare monoclonal antibodies against human thyroid peroxi-dase (hTPOmAb) and 29 hTPOmAbs had been developed by the classical hybridization technique. The relationship between thyroid microsome antibody (TMAb) and hTPOmAb was studied and it was found that the inhibition of TMAb to hTPOmAb binding with 125I-hTPO showed a dose-dependent relationship (r=0. 982,P<0. 001). By means of indirect immunofluorescence assay, it was also shown that hTPO only appeared on the membrane of human thyroid cell. These results demonstrate that the hTPOmAb developed are specific to human thyroid peroxidase.