抗HER2-hTNF-α新型免疫毒素的制备及功能研究  

Construction and Functional Study of a Novel Anti HER2/neu-hTNF-αImmunotoxin

作　　者：蒋琳[1] 冯静[2] 吴文芳[1] 杨东玲[2] 阎锡蕴[2] 

机构地区：[1]中国科学院沈阳应用生态研究所 [2]中国科学院生物物理研究所,北京100101

出　　处：《生物化学与生物物理进展》2005年第9期850-856,共7页Progress In Biochemistry and Biophysics

摘　　要：HER2/neu基因在肿瘤中的过度表达使其成为许多肿瘤的标志分子.为了增加过度表达HER2/neu的肿瘤细胞对肿瘤坏死因子(TNF)的敏感性和提高HER2/neu抗体的肿瘤杀伤效应,将抗HER2/neu单链抗体C6.5与人肿瘤坏死因子hTNF-α融合,构建了scFvC6.5-hTNF-α融合蛋白,完成了重组蛋白在大肠杆菌中的表达,产率为400μg/L菌液.经过亲和层析和柱复性,融合蛋白的纯度达95%以上.ELISA试验表明,scFvC6.5-hTNF-α能够特异结合HER2/neu阳性卵巢癌细胞SKOV-3和乳腺癌细胞MCF-7,而不结合HER2/neu阴性的黑色素瘤细胞A375.MTT试验表明,scFvC6.5-hTNF-α能够选择性地杀伤SKOV-3和MCF-7细胞,而不影响A375细胞的生长.这种肿瘤细胞特异性杀伤作用提示该免疫毒素具有肿瘤靶向治疗的潜在应用价值.HER2/neu is an attractive target for tumor therapy since its overexpression in a number of tumors. In order to enhance the cytotoxicity of anti-HER2/neu antibody, and the specificity of TNF-α for tumor cell, a fusion gene of antiHER2-hTNF-α was constructed. The recombinant fusion protein was produced in E.coli, and their refolding has done through affinity chromatography on a His-column. The ELISA assay showed that antiHER2-hTNF-α specifically bound to HER2/neu expressing SKOV-3 and MCF-7 cells, but it did not recognize the HER2/neu negative cells A375. In vitro study, it was found that antiHER2-hTNF-α inhibited the proliferation of SKOV-3 and MCF-7 cells, whereas it did not effect on the A375. These studies suggest that the recombinant antiHER2-hTNF-α immunotoxin has a potential application in HER2/neu expressing tumor therapy.

关 键 词：抗HER2/neu单链抗体 人肿瘤坏死因子 免疫毒素 

分 类 号：Q819[生物学—生物工程]