促甲状腺激素单克隆抗体的制备和应用

Preparation and Characterization of Monoclonal Antibodies Against Thyroid Stimulating Hormone (TSH)

目的:获得高效价、高特异性的促甲状腺激素(TSH)单克隆抗体,为建立高灵敏度的促甲状腺激素水平的测定方法奠定基础.方法:采用细胞融合技术,以TSH抗原免疫BALB/c小鼠的脾细胞与SP2/0骨髓瘤细胞进行融合.通过多次阳性筛选和有限稀释,建立了抗人TSH单克隆抗体杂交瘤细胞株.并对所建立的单抗进行了鉴定.结果:细胞融合率达100%,阳性率15%.共建立了5株杂交瘤细胞株,均属IgG1类.且与FSH、HCG、LH无交叉反应.McAb腹水效价在1:5×105～1:1×107.6个月液氮冻存后复苏培养测定细胞株的稳定性好.结论:本实验的融合率高,阳性率较高,所建立的细胞株的特异性高,McAb腹水效价高.为提高TSH检测的灵敏度和简便性提供了重要条件.

Objective To prepare quality monoclonal antibodies against thyroid stimulating hormone for highly sensitive assay. Methods BALB/c mice were immunized with synthetic TSH polypeptide. Spleen cells from the immunized mice were fused with myeloma cells of sp2/0, and cultured to produce hybridomas producing anti-human TSH monoclonal antibody. Results During cell fusion and screening, the rate of fusion was 100% and the psoitive rate of antibody producing was 15%. Five murine hybridomas producing monoclonal antibodies against TSH were estabilshed. All McAbs were of IgG 1 subclass and showed no cross reactivity with FSH, HCG and LH. The titers of the McAb ascitic fluid varied from 1∶5×10 5~1∶1×10 7. All murine hybridomas retained great stability after the freezing-thawing process. Conclusion The rate of cell fusion was high and the rate of positive antibody was good. Five species-specific murine hybridomas were established, producing McAbs of high titer. The McAbs could be excellent components in TSH assay with high sensitivity and simplicity.