| 抗Her-2-scFv-SEC2融合免疫毒素的构建和功能研究 |
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| 引用本文： | 徐明恺,张成刚,张惠文,周亚凤,张先恩,刘丽.抗Her-2-scFv-SEC2融合免疫毒素的构建和功能研究[J].生物化学与生物物理进展,2006,33(8):781-788. |
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| 作者姓名： | 徐明恺 张成刚 张惠文 周亚凤 张先恩 刘丽 |
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| 作者单位： | 1. 中国科学院沈阳应用生态研究所,沈阳,110016
2. 中国科学院武汉病毒研究所,武汉,430071 |
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| 摘 要： | 用基因工程方法,将金黄色葡萄球菌肠毒素 C2 与抗人表皮生长因子受体 HER-2 单链抗体 scFv-B1,以一连接短肽连接,构建融合免疫毒素 B-L-SEC2,并用改进的新型表达载体 pASK75-EX,在大肠杆菌 BL21(ED3)中表达. 以不溶性包涵体形式表达的目的蛋白经变性后以镍离子螯和层析纯化,并以透析法进行复性. 流式细胞术和 MTT 实验结果表明,纯化复性的融合免疫毒素 B-L-SEC2,在体外具有与 HER-2 过表达的靶细胞 SK-Br-3 特异性结合的活性,并对该细胞产生显著的特异性生长抑制作用.
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| 关 键 词： | 金黄色葡萄球菌肠毒素 C2 抗人表皮生长因子受体 HER-2 融合免疫毒素 肿瘤靶向 pASK75 表达载体 |
| 收稿时间： | 3/2/2006 8:01:05 PM |
| 修稿时间： | 5/15/2006 9:11:15 AM |
Fusion Immunotoxin Anti-HER-2-scFv-SEC2 Expressed in E. coli With an Improved Expression Vector pASK75-EX: Its Construction and Functioning |
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| XU Ming-Kai,ZHANG Cheng-Gang,ZHANG Hui-Wen,ZHOU Ya-Feng,ZHANG Xian-En and LIU Li.Fusion Immunotoxin Anti-HER-2-scFv-SEC2 Expressed in E. coli With an Improved Expression Vector pASK75-EX: Its Construction and Functioning[J].Progress In Biochemistry and Biophysics,2006,33(8):781-788. |
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| Authors: | XU Ming-Kai ZHANG Cheng-Gang ZHANG Hui-Wen ZHOU Ya-Feng ZHANG Xian-En and LIU Li |
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| Institution: | Institute of Applied Ecology, The Chinese Academy of Sciences, Shenyang 110016, China;Institute of Applied Ecology, The Chinese Academy of Sciences, Shenyang 110016, China;Institute of Applied Ecology, The Chinese Academy of Sciences, Shenyang 110016, China;Wuhan Institute of Virology, The Chinese Academy of Sciences, Wuhan 430071, China;Wuhan Institute of Virology, The Chinese Academy of Sciences, Wuhan 430071, China;Institute of Applied Ecology, The Chinese Academy of Sciences, Shenyang 110016, China |
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| Abstract: | A tumor-targeting recombinant fusion immunotoxin B-L-SEC2 was constructed by fusing staphylococcal enterotoxin C2(SEC2)and an anti-HER-2 single-chain Fv B1 through a peptide linker,and expressed in E.coli strain BL21(DE3)with an improved expression vector pASK75-EX as inclusion body.The denatured inclusion body was purified with Ni-NTA chelate agarose,and then re-natured by dialysis.FACS and MTT assays indicated that the re-natured fusion immunotoxin B-L-SEC2 could target the HER-2 over-expressing breast tumor cell SK-Br-3 in vitro,and inhibit the growth of SK-Br-3. |
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| Keywords: | staphylococcal enterotoxin C2(SEC2) HER-2 fusion immunotoxin tumor-targeting pASK75 |
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