|
|||||
|
|
| 可内化的人源抗Met基因工程抗体Fab的亲和力成熟与特性分析 | |
| 引用本文： | 朱 进,赵 萍,焦永军,王 昕,曹伯良,冯振卿,管晓虹.可内化的人源抗Met基因工程抗体Fab的亲和力成熟与特性分析[J].生物化学与生物物理进展,2007,34(1):73-79. |
| 作者姓名： | 朱 进 赵 萍 焦永军 王 昕 曹伯良 冯振卿 管晓虹 |
| 作者单位： | 1. 南京医科大学卫生部抗体技术重点实验室,南京,210029;南京军区军事医学研究所,南京,210002 2. 南京医科大学卫生部抗体技术重点实验室,南京,210029;Van Andel Research Institute,Antibody,Laboratory MI,49503,USA 3. 南京医科大学卫生部抗体技术重点实验室,南京,210029 |
| 摘 要： | 应用噬菌体展示技术制备抗Met(HGF受体,一个与肿瘤发生、侵袭和转移相关原癌基因产物)特异性、高亲和力的全人Fab片段.Fab基因分三步合成,以从错配PCR突变库中筛选出的Fab基因可变区为模板,扩增VH和VL基因,分别与CH1、CL基因融合,合成Fd和L基因,再拼接合成Fab基因,克隆于pComb3XSS中,构建Fab次级抗体库.经细胞筛选和固相筛选,获得高亲和力阳性克隆.工程菌经IPTG诱导表达,SDS-PAGE和蛋白质印迹分析,在25ku和27ku出现预期大小蛋白质条带.Fab分子经流式细胞术、免疫沉淀、细胞免疫荧光检测,结果表明,Fab能够与S114和MKN45细胞膜上的Met胞外区特异性结合,而与阴性细胞NIH3T3不结合.抗体内化分析显示,Fab能够与标记肥皂草毒素(ZAP)的抗人IgG结合,并进入细胞内,抑制Met阳性细胞的生长,揭示该抗体能够与Met特异性结合,并且被细胞内化.该抗体有望成为肿瘤临床诊断或治疗的候选分子. |
| 关 键 词： | 肝细胞生长因子受体 噬菌体抗体库 亲和力成熟 内化抗体 |
| 收稿时间： | 2006/5/31 0:00:00 |
| 修稿时间： | 2006-05-31 |
Affinity Maturation and Characterization of Internalized Human Anti-Met Recombinant Antibody Fab |
|
| ZHU Jin,ZHAO Ping,JIAO Yong-Jun,WANG Xin,CAO Bo-Liang,FENG Zhen-Qing and GUAN Xiao-Hong.Affinity Maturation and Characterization of Internalized Human Anti-Met Recombinant Antibody Fab[J].Progress In Biochemistry and Biophysics,2007,34(1):73-79. | |
| Authors: | ZHU Jin ZHAO Ping JIAO Yong-Jun WANG Xin CAO Bo-Liang FENG Zhen-Qing and GUAN Xiao-Hong |
| Institution: | Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, China;Huadong Medical Institute of Biotechniques Nanjing 210002, China;Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, China;Van Andel Research Institute, Antibody Laboratory, MI 49503, USA;Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, China;Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, China;Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, China;Van Andel Research Institute, Antibody Laboratory, MI 49503, USA;Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, China;Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, China |
| Abstract: | |
| Keywords: | hepatocyte growth factor receptor (Met) phage-displayed antibody library affinity maturation internalized antibody |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |
| 点击此处可从《生物化学与生物物理进展》浏览原始摘要信息 | |
| 点击此处可从《生物化学与生物物理进展》下载免费的PDF全文 | |