人精子膜结合腺苷脱氨酶单克隆抗体的制备和初步鉴定

The preparation of anti human sperm plasma membrane bound adenosine deamianse monoclonal antibody and its preliminary characterization

本研究以正常人精子，用ｐｅｒｃｏｌｌ分离，经脾直接免疫、细胞融合和克隆化后，以标准ＡＤＡ为抗原筛选，得到２８株单克隆抗体（ＭｃＡｂ）细胞株。其腹水效价均在１：６４００以上（ＥＬＩＳＡ）。免疫球蛋白分类显示，１株为ＩｇＭ，其余各株皆为ＩｇＧ。ＭｃＡｂ对ＡＤＡ活性的抑制实验结果显示，有２２株ＭｃＡｂ对ＡＤＡ有抑制作用而其余６株对ＡＤＡ无抑制作用。初步研究结果显示我们得到的ＭｃＡｂ具有特异性，可用于对精子中ＡＤＡ的性质研究。

bjective :To prepare a specific monolconal antibody against human sperm plasma membrane bound adenosine deaminase.Mothods : Hybridoma techniques.esults:After direct injection of well washed intact normal human sperm into 4Balb/c mice spleen for 90 hours.The splenic cells were fusioned with SP210 cells.Twenty- six monoclonal antibodies were obtained after the screening with purified adenosine deaminase as the antigen.The range of titers for these antibodies were from 1 :6 400 to 1 : 5 0 000 ( ELISA).Six monoclonal antibodies were found to have no effect on adenosine deaminase activity ,but rest of them do.Conclusions :Direct immunization is a useful method for preparing some expensive antigen′s monoclonal antibody,and is a time saving assay as well. Our preliminary study indicates that the monoclonal antibodies we prepared are specific for the sperm plasma membrane found adenosine deaminase.To characterizie the adenosine deaminase in sperm with it monoclonal antibodies is obviously our new focus in further .