人卵泡刺激素β亚基单克隆抗体的制备及鉴定

Preparation and identification of monoclonal antibody against human follicle stimulating hormone beta subunit

目的 建立分泌抗人卵泡刺激素 (FSH) β亚基 5 1～ 6 0片段单克隆抗体细胞株 ,并对单抗进行免疫学鉴定。 方法 以人工合成的FSHβ蛋白抗原决定簇多肽为免疫原 ,采用脾内包埋法免疫Balb c小鼠 ,取其脾细胞与SP2 0小鼠骨髓瘤细胞融合 ,杂交瘤细胞用人工合成的人FSHβ亚基 5 1～ 6 0片段包被、间接ELISA法筛选。结果 获得稳定分泌抗人FSHβ片段单抗细胞 1株 (F1C)。ELISA检测腹水效价为 1∶1.6× 10 4 ,与黄体生成素 (LH)、促甲状腺素 (TSH)、人绒毛膜促性腺激素 (hCG)等无交叉反应 ,免疫印迹 (Westernblot)显示该单抗特异性强。结论 本实验所制备的单抗具有良好的灵敏性、特异性和实用性 ,可用于酶免疫检测方法的建立 。

Objective To establish anti human follicle stimulating hormone(FSH) beta subunit 51-60 fragment antibody cell line, and to perform immunological identification to the antibody. Method Balb/c mice were immunized by embedding the nitrocellulose paper strips into the spleen dipped in purified artificial synthetic amino acids(51-60) region of FSH β protein buffer. The splenic cell of the mice fused with SP2/0 mouse myeloma cells. The hybridoma culture supernants were screened by indirect ELISA with FSH β fragment as antigen. Results One hybridoma cell line against FSH β McAb was successfully obtained. The titers of ascites were 1∶1.6×10 4 and the McAb had no cross reactions with LH,TSH and hCG. Western blot analysis showed that the antibody can bind with FSHβ specifically. Conclusion The McAb has good specificity, reactivity and usefulness, which can be used for enzyme linked immunoassay and a ELISA diagnostic kit for clinical detection.