【摘要】 采集６～８周人工流产胎盘绒毛，经胶原酶处理及梯度离心后得到滋养细胞，再用ＮＰ－４０提取滋养细胞抗原。经常规免疫、细胞融合和克隆化后得到２０多株单克隆抗体（ＭｃＡｂ）细胞株。其中１０株ＭｃＡｂ进行了免疫学鉴定。免疫球蛋白分类，２株为ＩｇＭ，８株为ＩｇＧ，其中４株为ＩｇＧｌ。免疫斑点试验结果显示，１０株ＭｃＡｂ均为阳性，其滴度为１：８０～１：１２８０。免疫荧光定位，１０株中７株与滋养层细胞反应，２株与绒毛基底膜结合，１株为阴性。并对其进行了免疫转移印迹试验和交叉反应性研究，初步研究结果显示，单抗Ｔ６有着较好的特异性，值得进一步研究。更多还原

【Abstract】 The placenta villi of 6～8weeks of gestation were collected from normal pregnant womenafter induced abortion The trophoblast cells were obtained by using collagenase treatment and gradient contrifugation. The trophoblast antigens were prepared by iising NP-40 extraction method.After routineimmunization of BALB/c mice and cell fusion,the hvbridoma cells were established.More than twentycell clones secreting monoclonal antibodies(McAbs)were obtained by using clonal selection technique.The immunofluorescent localization assay,Western blot assay, Bio-dot assay,immunoglobulin type ana-lysis and crossreactivity study were performed in the present study for identification of these McAbs.Theresults of immunoglobulin type analysis showed that 2 McAbs were lgM,8 McAbs were IgG, amongthem 4 McAbs were IgGl With Bio-dot assay,Ten McAbs tested all had positive reaction withtrophoblast antigens,their titres ranged from 1 :80 to 1:1 28O. In immunofluorescent locaiization assay,7/10 McAbs reacted with trophoblast cellS, and the other two reacted with basement membrane ofchorionic villi. There was negative fluorescent reaction only in one The molecular weights of trophoblastantigens recognized by tiiese McAbs were 145 000, 115 000,110 000, 97 000,80 000,74 000, 70 000 and44 000 All McAbs exhibited negative reactions with hCG,While 3 / 1 0 and 1 / 10 McAbs had positivereaction with lymphocyte cells and red blood cells,respectively The preliminary results showed that T6McAb was more specific to trophoblast antigen. Further studies of these McAb need to be continued.更多还原