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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。
Objective To develop anti-human FKBP5 2 monoclonal antibody (mAb) and analyze its immunobiological properties. Methods BALBc mice were immunized with purified human FKBP5 2 protein. The specificity of the mAb was identified by ELISA. The relative molecular mass (Mr) and the functional region of human FKBP5 2 molecule recognized by the 9 mAbs were identified by Western blot. Results Nine hybridoma cells secreting anti-human FKBP5 2mAb were obtained. The anti-human FKBP5 2 mAb recognizes human FK BP5 2 protein expressed in a prokaryotic system with a relative Mr of 5200 and also recognizes a native FKBP5 2 protein of Mr 52 000 in the cytoplasm of Jurkat cells. Both mAbs recognize the second functional region in human FKBP5 2 molecule. The remaining seven mAbs recognize the third functional region in human FKBP5 2 molecule. Conclusion The anti-human FKBP5 2 mAb was successfully prepared.