目的: 制备分泌特异性抗人甲状腺球蛋白(thyroglobulin,Tg)单克隆抗体的杂交瘤细胞株, 为建立高灵敏度的Tg检测方法做准备。方法: 以天然人源甲状腺球蛋白为抗原经皮下免疫BALB/c小鼠, 通过细胞融合制备分泌抗人甲状腺球蛋白单克隆抗体, 并对其进行特异性鉴定, 建立检测Tg的双抗体ELISA(enzyme-linked immunosorbent assay)夹心法。结果: 获得7株可稳定分泌抗人甲状腺球蛋白单克隆抗体的杂交瘤细胞株, 经ELISA鉴定, 筛选抗体可与Tg抗原有良好的特异性反应。建立的双抗体夹心ELISA方法敏感性可达1 ng/mL。 结论: 成功制备了抗人Tg单克隆抗体并建立了检测人Tg双抗体夹心ELISA方法, 为进一步研发Tg快速诊断试剂盒提供了原料。

Objective: To prepare monoclonal antibody against human thyroglobulin and establish a highly sensitive method to detect human thyroglobulin. Methods: BALB/c mice were subcutaneously immunized by native human thyroglobulin, the splenocytes of immunized mice were fused with SP2/0 myeloma cells by a routine method. The hybridoma cells secreting specific antibody were detected. The stability of the obtained hybridoma cells and the specificity of anti-thyroglobulin monoclonal antibody the hybridoma cells secreted were identified. Based on all above, the double antibody sandwich ELISA method was established to detect thyroglobulin. Results: Seven hybridoma cell lines which could secrete monoclonal antibody stably were obtained, and ELISA indicated the antibodies had a good specificity and sensitivity to Tg antigen. A sandwich ELISA kit for detecting human thyroglobulin was established, and its sensitivity reached to 1 ng/mL. Conclusion: The anti-thyroglobulin monoclonal antibodies were prepared and a sandwich ELISA kit for the detection of human thyroglobulin was established successfully, which would provide a raw material for the development of Tg rapid diagnostic kits.