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Her2全人源抗体的制备及生物学功能研究
Construction and Characterization of Fully Human Anti-Her2 Antibodies
【作者】 方晨；
【导师】 郭亚军；
【作者基本信息】 第二军医大学 ， 肿瘤学， 2011， 博士
【摘要】 目的：Her2是一种具有酪氨酸激酶活性的跨膜糖蛋白,它的过表达与肿瘤形成、恶性程度及预后密切相关。本研究的目的是通过构建全人源大容量噬菌体抗体库从而筛选鉴定具有高效抗肿瘤活性的Her2全人源抗体。方法：本实验构建了全人源大容量噬菌体抗体库,表达纯化了人Her2膜外区蛋白(Her2ECD),并利用Her2 ECD对噬菌体抗体库进行筛选,获得了异性结合Her2的噬菌体单链抗体ScFv,并构建完整L/pcDNA3.1(+)和H/pcDNA3.1(+)表达载体,在真核细胞CHO细胞中成功地表达了抗Her2抗体并对它们的体外抗肿瘤作用进行了研究。结果：我们制备的Her2抗体在体外能有效抑制乳腺癌细胞的MAPK、Akt信号通路,抑制Her2高表达乳腺癌细胞系SK-BR-3和BT-474的生长。结论：通过噬菌体抗体库的筛选,我们获得了具有抗肿瘤活性的Her2全人源抗体。
【Abstract】 Objective:Her2 is a tyrosine kinase activity transmembrane glycoprotein, and its overexpression has relationship with tumor formation, the degree of malignancy and prognosis. In this study, we aim to prepare fully human anti-HER2 antibodies with potent antitumor activity. Methods:We constructed a large-capacity human ScFv phage antibody library and selected ScFv specific for HER2. Then the antibody expression vectors were constructed and anti-Her2 antibodies were expressed in CHO cells. Finally, we investigated the in vitro antitumor effects of these anti-Her2 antibodies on breast cancer cell lines. Results:Anti-Her2 antibodies could effectively inhibit the growth of Her2 high expresstion breast cancer cell lines SK-BR-3 and BT-474 in vitro. Conclusion:The present study provides several fully human anti-Her2 antibodies with potent antitumor activity and these anti-Her2 antibodies may serve as potential therapeutic agents for breast cancer.