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目的:制备抗结肠肿瘤相关抗原的单克隆抗体(mAb)并对其进行初步的鉴定及功能研究。方法:将结肠癌细胞系或磨碎的结肠癌组织免疫小鼠,按常规方法进行细胞融合,采用活细胞周围花环形成的方法进行阳性克隆的筛选。用间接免疫荧光染色和流式细胞术分析及酶免疫组织化学的方法进行初步鉴定,并用抗体介导的补体依赖的细胞毒试验(CDC)进行初步的功能研究。结果:获得了2株分泌抗结肠肿瘤相关抗原mAb杂交瘤细胞株3A12和6A8。流式细胞仪检测,2株杂交瘤分泌的抗体均能在结肠癌细胞系中均为阳性。普通石蜡切片及组织芯片的免疫酶组织化学染色结果,显示2株mAb均在结肠肿瘤组织中有较高的阳性率。2株mAb分别与靶细胞及补体作用后,对于靶细胞的杀伤率分别为98.1%和42.4%。结论:成功地制备了2株识别结肠癌细胞表面抗原的mAb,在结肠癌的诊断和治疗中可能具有潜在的应用价值。
OBJECTIVE: To prepare monoclonal antibody (mAb) against colon tumor associated antigen and conduct preliminary identification and functional studies. Methods: The colon cancer cell lines or ground colon cancer tissues were immunized with the mice, and the cells were fused according to a conventional method. The positive clones were screened by the method of forming the wreath around the living cells. Indirect immunofluorescence staining and flow cytometry analysis and enzyme immunohistochemistry were used for preliminary identification and preliminary functional studies were performed using antibody-mediated complement-dependent cytotoxicity assay (CDC). Results: Two strains of McAbs 3A12 and 6A8 secreting anti-colon cancer-associated mAb were obtained. Flow cytometry, two hybridoma secreted antibodies in colon cancer cell lines were positive. Immunohistochemical staining results of ordinary paraffin sections and tissue microarrays showed that both mAbs had a higher positive rate in colon tumor tissues. The killing rates of the two mAbs against the target cells were respectively 98.1% and 42.4% with the target cells and the complement respectively. Conclusion: Two mAbs that recognize the surface antigens of colon cancer cells were successfully prepared, which may have potential value in the diagnosis and treatment of colon cancer.