抗人FKBP52分子单克隆抗体的制备和鉴定

Preparation and characterization of monoclonal antibodies to human FKBP52 molecule

目的 研制抗人FKBP5 2单克隆抗体 (mAb) ,并分析其免疫生物学特性。方法采用纯化的人FKBP5 2蛋白免疫BALB c小鼠。用ELISA法鉴定mAb的特异性 ;用Westernblot鉴定制备的 9株mAb所识别的抗原相对分子质量 (Mr)及结合人FKBP5 2分子的功能区。结果共获得9株分泌抗人FKBP5 2mAb的杂交瘤细胞。抗人FKBP5 2mAb可识别相对Mr为 5 2 0 0 0在原核系统中表达的人FK BP5 2蛋白 ,也可识别Jurkat细胞浆中Mr为 5 2 0 0 0的天然FKBP5 2蛋白。两株mAb均可识别人FKBP5 2分子中的第2功能区。其余 7株mAb可识别人FKBP5 2分子中的第 3功能区。结论成功地制备了抗人FKBP5 2分子mAb。

Aim To prepare monoclonal antibodies(mAb) to human FKBP52 molecule and analyse their biological properties. Methods BALB/c mice were immunized with hFKBP52 protein purified from prokaryotic expression system. Specificity and antigen-binding sites of the mAbs were determined by ELISA and Western blot, respectively. Results Nine hybridoma cell lines that could secret anti-FKBP52 mAb were established. The mAbs bound mainly to natural FKBP52 protein with Mr 52 000 molecule in cytoplasma of Jurkat cells. Two of 9 hybridoma cells could recognize the second domain of hFKBP52, the rest recognized the third domaine. Conclusion The mAbs to human FKBP52 molecule have been prepared and characterized successfully.