抗肝癌血管内皮细胞的单克隆抗体的制备和鉴定

目的　研究制备抗肝癌区血管内皮细胞的单克隆抗体，并进行初步的鉴定。方法　单克隆抗体制备：将制备好的抗原、免疫动物，行ＰＥＧ 细胞融合、免疫组织化学和ＥＬＩＳＡ抗体筛选，进行细胞克隆。抗体的生产及纯化；组织学鉴定分析：亲和层析蛋白分离、ＳＤＳＰＡＧＥ 电泳、免疫组织化学分析；抗体的体外细胞毒作用试验选用补体介导的细胞毒实验ＭＴＴ法。结果　以经肝癌细胞系培养上清诱导扩增的人胎脐静脉血管内皮细胞为抗原，制备并获得了抗肝癌区血管内皮细胞的单克隆抗体；组织学研究表明，抗体在肝癌组织的血管内皮细胞中有特异性，在体外对诱导扩增的血管内皮细胞有补体介导的细胞毒作用。结论　制备并获得抗肝癌区血管内皮细胞的单克隆抗体；抗体在体外具有细胞毒作用。

Preparation and identification of monoclonal antibodies against vascular endothelial cells in hepatocarcinoma site

Objective To prepare and identify monoclonal antibodies against vascular endothelial cells in hepatocarcinoma. Methods Cell culture, animal immunization, PEG cell merging, cell cloning, immunohistochemical sABC method and enzyme linked immunosorbent assay (ELISA), SDS PAGE electrophoresis, affinity chromatography, in vitro cytotoxic effect of the monoclonal antibodies, cell culture, and MTT method were performed. Results The monoclonal antibodies against vascular endothelial cells in hepatocarcinoma area were prepared using fetal umbilical vein endothelial cells as an immunogen and were amplified after induction by supernatant of cultured hepatocarcinoma cell lines. The antibodies had high specificity and showed cytotoxic effect on amplifying vascular endothelial cells induced in vitro. Conclusion Monoclonal antibody against vascular endothelial cells can be made in hepatocarcinoma site and has cytotoxic effect in vitro.