抗人滋养层细胞表面抗原-2单抗的制备及免疫学特性分析

Preparation and immunologic characterization of a monoclonal antibody against human trophoblast cell-surface antigen 2

目的:利用杂交瘤技术制备抗人滋养层细胞表面抗原-2(trophoblast cell-surface antigen 2,Trop2)单克隆抗体并鉴定其免疫学特性。方法:以胰腺癌细胞系BxPC3免疫BALB/c小鼠,取其脾细胞与SP2/0细胞融合后制备单抗。通过酶联免疫法、免疫荧光、免疫沉淀和质谱分析、流式细胞术、免疫组织化学等方法,鉴定单抗的免疫学特性。结果:通过细胞的融合与筛选,获得了持续分泌抗Trop2单抗的杂交瘤细胞株,该抗体可以识别细胞表面的Trop2膜蛋白,也可用于免疫组化识别人肿瘤组织中的Trop2蛋白,同时该抗体对乳腺癌细胞的生长具有一定的抑制作用。结论:该单抗具有作为Trop2阳性表达肿瘤靶向治疗的潜在应用价值。

Objective：To produce a monoclonal antibody（mAb） against human trophoblast cell-surface antigen 2（Trop2） with hybridoma technology and identify its immune specificity.Methods：The spleen cells from BALB/c mice immunized with pancreatic cancer cell line BxPC3 cells were fused with SP2/0 cells to produce a monoclonal antibody against human Trop2.Further,several immunological methods such as ELISA,IF,IP,MALDI-TOF MS,FACS and IHC were used to characterize the immune specificity of the mAb.Results：The results demonstrated that the specific mouse anti-human Trop2 mAb was successfully produced and can be secreted continuously and steadily.Moreover,this mAb can effectively recognize the transmembrane glycoprotein Trop2 over-expressed in human cancer cells and tissues.Conclusion：We successfully obtained mouse anti-human Trop2 mAb which may be potentially utilized for some tumor targeted therapy.