人肺癌单克隆抗体的制备及其抗原分子的鉴定

Preparation of Monoclonal Antibody Against Lung Cancer And Identification of Its Targeting Antigen

用人肺癌细胞株A549的细胞膜蛋白作为免疫原,采用杂交瘤技术成功制备了抗人肺癌细胞单克隆抗体McAb4E7,并利用双向电泳、免疫印迹杂交、肽指纹图谱和MALDI-TOF-MS串连质谱分析鉴定其相关抗原分子,发现其相关抗原在人肺癌细胞株A549及人肝癌细胞株HepG2中有表达,且在A549细胞中表达量较高。最后得到一个在A549细胞中表达的与单克隆抗体McAb4E7特异性结合的蛋白,并鉴定出这个蛋白是ATP合成酶β亚基,从而证明了ATP合成酶β亚基就是肺癌单克隆抗体McAb4E7的靶抗原,其可作为肺癌具有潜在治疗意义的靶点。

A mouse-anti-huamn monoclonal antibody was produced by using the membrane proteins of human lung carcinoma cell line A549 as the immunogen to generate monoclonal antibodies against lung carcinoma with the use of hybridoma techniques.McAb4E7 was prepared successfully.To identify its antigen,proteomic technologies such as two-dimenstional electrophoresis,western blotting and mass spectrometry were employed.The targeting antigen of McAb4E7 expressed positive in human lung cancer cell lines A549 and human hepatocarcinoma cell line HepG2,moreover,the expression of the antigen was stronger in A549 cells.Finally,we obtained one positive protein in A549 cell line that has strong affinity and specificity for McAb4E7,which was identified to be ATP synthase β subunit.We identified ATP synthase β subunit as the targeting antigen of lung carcinoma special monoclonal antibody McAb4E7.