抗人F1-F0 ATP合成酶beta亚基单抗的制备及其抗肿瘤活性研究

被引:8
作者
张霞 [1 ]
彭艳 [2 ]
俞丽丽 [1 ]
陈建鹤 [2 ]
王怡波 [3 ]
林建波 [3 ]
倪健 [2 ]
殷明 [1 ]
机构
[1] 上海交通大学药学院
[2] 苏州工业园区晨健抗体组药物开发有限公司
[3] 第二军医大学肝胆外科医院综合治疗一科
关键词
F1-F0ATP合成酶beta亚基; 单克隆抗体; 多柔比星; 肿瘤耐药;
D O I
暂无
中图分类号
R73-36 [治疗实验];
学科分类号
100214 ;
摘要
目的:制备鼠抗人F1-F0ATP合成酶beta亚基(hATP5B)单抗,并对其特异性和抗肿瘤活性进行研究。方法:以原核表达人hATP5B免疫BALB/c小鼠,通过杂交瘤技术,筛选分泌抗hATP5B单抗的杂交瘤细胞株。采用Protein A亲和层析法纯化抗体腹水,SDS-PAGE检测纯化产物纯度。利用Western blot、细胞免疫荧光对其特异性进行鉴定,并通过抑制乳腺癌细胞MCF-7及其耐药株MCF-7/Adr表面ATP生成,细胞毒性实验进行抗肿瘤活性研究。结果:获得一株稳定表达抗hATP5B单抗的杂交瘤细胞株Mab3B8,Western blot和细胞免疫荧光结果表明,该抗体与乳腺癌MCF-7及其耐药细胞MCF-7/Adr细胞天然抗原结合;可明显抑制MCF-7及其耐药株MCF-7/Adr细胞膜表面ATP合成;与化疗药物多柔比星(曾用名:阿霉素)联合作用,可降低化疗药物多柔比星对肿瘤细胞的IC50。结论:成功建立了一株可特异性识别天然hATP5B的单抗,有阻断肿瘤细胞膜表面ATP合成活性并可明显增强MCF-7及其耐药株MCF-7/Adr细胞对多柔比星敏感性的作用。此项研究对膜表达F1-F0ATP合成酶功能探讨及肿瘤治疗研究都具有重要意义。
引用
收藏
页码:984 / 987+992 +992
页数:5
相关论文
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