| 人源抗肝癌单链抗体SA3与增强型绿色荧光蛋白的融合表达及其体内靶向性研究(英文) |
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| 引用本文： | 黄建,李跃辉,郭锋杰,童永清,王甲甲,胡锦跃,李官成.人源抗肝癌单链抗体SA3与增强型绿色荧光蛋白的融合表达及其体内靶向性研究(英文)[J].中南大学学报(医学版),2011,36(10). |
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| 作者姓名： | 黄建 李跃辉 郭锋杰 童永清 王甲甲 胡锦跃 李官成 |
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| 作者单位： | 中南大学肿瘤研究所,卫生部癌变原理重点实验室,教育部癌变与侵袭原理重点实验室,长沙410078 |
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| 基金项目： | supported by the National Natural Science Foundation of China(NSFC)-973 Project(2010CB833605); the Science&Technology Department Research Fundation of Hunan Provience(2010SK3132); the Health Department Research Fundation of Hunan Provience(A2003001),P.R China |
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| 摘 要： | 目的:探讨人源抗肝癌单链抗体SA3与增强型绿色荧光蛋白(EGFP-SA3)的融合表达、纯化、复性及其体内靶向性。方法:将SA3,EGFP基因,插入pET-25b(+),构建EGFP-SA3/pET-25b(+)原核表达载体,测序鉴定后转化大肠杆菌BL21(DE3);IPTG诱导融合蛋白EGFP-SA3的表达,复性、纯化后经SDS-PAGE鉴定;EGFP-SA3与HepG2细胞经体外孵育后在荧光显微镜下观察单链抗体SA3与肝癌细胞的结合作用;然后通过尾静脉将其注射入荷肝癌裸鼠体内观察EGFP-SA3的体内靶向作用。结果:SA3,EGFP基因分别插入载体pET-25b(+)后,重组表达载体EGFP-SA3/pET-25b(+)分别行NcoI-XhoI和NcoI-EcoRI双酶切,结果发现在750 bp左右出现一条带,与EGFP基因大小一致,在1.5 kb左右处出现一条带,与EGFP和SA3两个基因片段的总大小一致,DNA测序结果证实融合表达载体EGFP-SA3/pET-25b(+)构建成功;重组表达载体EGFP-SA3/pET-25b(+)经IPTG诱导表达后,SDS-PAGE显示融合蛋白EGFP-SA3分子质量...
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| 关 键 词： | 肝癌 scFv EGFP 融合蛋白 靶向治疗 原核表达 |
Expression of scFv SA3 against hepatoma fused with enhanced green fluorecsent protein and its targeted ability in vivo |
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| HUANG Jian,LI Yuehui,GUO Fengjie,TONG Yongqing,WANG Jiajia,HU Jinyue,LI Guancheng.Expression of scFv SA3 against hepatoma fused with enhanced green fluorecsent protein and its targeted ability in vivo[J].Journal of Central South University (Medical Sciences)Journal of Central South University (Medical Sciences),2011,36(10). |
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| Authors: | HUANG Jian LI Yuehui GUO Fengjie TONG Yongqing WANG Jiajia HU Jinyue LI Guancheng |
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| Institution: | HUANG Jian,LI Yuehui,GUO Fengjie,TONG Yongqing,WANG Jiajia,HU Jinyue,LI Guancheng(Cancer Research Institution,Central South University,Key Laboratory of Carcinogenesis,Ministry of Health,Key Laboratory of Carcinogenesis and Invasion,Ministry of Education,Changsha 410078,China) |
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| Abstract: | Objective To express and purify the human scFv antibody,SA3,against the hepatoma fused to enhanced green fluorecsent protein,and to observe the targeted capacity of fusion protein EGFP-SA3 in vivo.Methods SA3 and EGFP genes were cloned into plasmid pET-25b(+) to construct the recombinant plasmid EGFP-SA3/pET-25b(+),followed by DNA sequencing.Then it was transformed into E.coli BL21(DE3) and induced for fusion expression of EGFP-SA3 with IPTG.The expressed fusion protein EGFP-SA3 was purified and detected wi... |
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| Keywords: | hepatoma scFv EGFP fusion expression targeted therapy prokaryotic expression |
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