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| 人黑素瘤单链抗体-量子点纳米探针的制备及其与黑素瘤细胞特异性结合 | |
| 引用本文： | 张小敏,张堂德,鲍晨晨,宋 华,李 娜,刘 彬,贺 蓉,李智铭,崔大祥,任秋实.人黑素瘤单链抗体-量子点纳米探针的制备及其与黑素瘤细胞特异性结合[J].中国肿瘤生物治疗杂志,2010,17(1):30-35. |
| 作者姓名： | 张小敏 张堂德 鲍晨晨 宋 华 李 娜 刘 彬 贺 蓉 李智铭 崔大祥 任秋实 |
| 作者单位： | 1. 南方医科大学,珠江医院,皮肤科,广东,广州,510280 2. 上海交通大学,微纳科学研究院,生物纳米工程研究室,上海,200240 3. 上海交通大学,生命科学技术学院,上海,200240 |
| 基金项目： | 国家高技术研究发展（863）计划重点项目（No. 2007AA022004）；国家重点基础研究发展（973）计划资助项目（No. 2010CB933900） |
| 摘 要： | 目的：制备抗人黑素瘤细胞单链抗体与碲化镉量子点(CdTe guantun dot, CdTe)连接的纳米探针，观察其对恶性黑素瘤细胞的特异性结合效果。方法：将抗人黑素瘤神经节苷脂单链抗体（antihuman melanoma ganglioside single chain variable fragment antibody，GD/ScFvMEL）基因克隆到pET32a（+）载体中，然后在BL21（DE3）细菌中进行诱导表达，采用变性法纯化表达的蛋白，再用改良的透析法复性目的蛋白，SDSPAGE分析表达的GD/ScFvMEL抗体蛋白。制备的GD/ScFvMEL抗体与量子点连接制备成纳米探针GD/ScFvMELQDs，然后与黑素瘤A375细胞株及胃癌MGC803细胞株孵育，观察纳米探针与肿瘤细胞结合的特异性。结果：PCR、双酶切和序列测定证实重组子的拼接完全正确，SDSPADE结果显示，GD/ScFvMEL抗体的表达量达40%。纯化复性后的GD/ScFvMEL抗体连接量子点制备GD/ScFvMELQDs纳米探针，经扫描电镜、X衍射分析、荧光光谱和SDSPAGE分析证实纳米探针的成功制备。GD/ScFvMELQDs纳米探针能特异性结合A375黑素瘤细胞，不与胃癌细胞MGC803细胞结合。结论：成功制备抗人黑素瘤神经节苷脂单链抗体量子点纳米探针，该探针可特异性结合黑素瘤细胞。 |
| 关 键 词： | 黑素瘤 神经节苷脂 单链抗体 量子点 纳米探针 |
| 收稿时间： | 2009/11/5 0:00:00 |
| 修稿时间： | 2009/12/10 0:00:00 |
Preparation of human malignant melanoma ganglioside ScFv antibody conjugated quantum dot nanoprobe and its specific binding with human malignant melanoma cells |
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| ZHANG Xiao-min,ZHANG Tang-de,BAO Chen-chen,SONG Hua,LI Na,LIU Bin,HE Rong,LI Zhi-ming,CUI Da-xiang,REN Qiu-shi.Preparation of human malignant melanoma ganglioside ScFv antibody conjugated quantum dot nanoprobe and its specific binding with human malignant melanoma cells[J].Chinese Journal of Cancer Biotherapy,2010,17(1):30-35. | |
| Authors: | ZHANG Xiao-min ZHANG Tang-de BAO Chen-chen SONG Hua LI Na LIU Bin HE Rong LI Zhi-ming CUI Da-xiang REN Qiu-shi |
| Institution: | Department of Dermatology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong, China;Department of Dermatology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong, China;Department of Bio/Nano Science and Engineering, Research Institute of Micro/Nano Science and Technology, Shanghai Jiaotong University, Shanghai 200240, China;Department of Bio/Nano Science and Engineering, Research Institute of Micro/Nano Science and Technology, Shanghai Jiaotong University, Shanghai 200240, China;Department of Bio/Nano Science and Engineering, Research Institute of Micro/Nano Science and Technology, Shanghai Jiaotong University, Shanghai 200240, China;Department of Bio/Nano Science and Engineering, Research Institute of Micro/Nano Science and Technology, Shanghai Jiaotong University, Shanghai 200240, China;Department of Bio/Nano Science and Engineering, Research Institute of Micro/Nano Science and Technology, Shanghai Jiaotong University, Shanghai 200240, China;Department of Bio/Nano Science and Engineering, Research Institute of Micro/Nano Science and Technology, Shanghai Jiaotong University, Shanghai 200240, China;Department of Bio/Nano Science and Engineering, Research Institute of Micro/Nano Science and Technology, Shanghai Jiaotong University, Shanghai 200240, China;College of Life Science and Biotechnology, Shanghai Jiaotong University, Shanghai 200240, China |
| Abstract: | Objective:To prepare a nanoprobe,anti-human melanoma ganglioside single chain variable fragment (GD/ScFvMEL) antibody conjugated with CdTe quantum dot,and to observe its ability to specifically bind human malignant melanoma cells.Methods:The GD/ScFvMEL gene was cloned into pET32a (+),and the plasmid was then transformed into E.coli BL21 (DE3) for GD/ScFvMEL protein antibody expression.The expressed GD/ScFvMEL antibody was purified by denaturing method and further refolded by modified dialysis method.The pur... |
| Keywords: | melanoma gaglioside single-chain antibody quantum dot nanoprobe |
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