| 结核分枝杆菌Rv0733 6His抗原羊驼单域重链抗体的筛选与鉴定 |
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| 引用本文： | 潘欣,蔡家麟,王颖,陈建霞,曾思良,方伟,陈敏,廖万清,余秀珍,张俊,宋维.结核分枝杆菌Rv0733 6His抗原羊驼单域重链抗体的筛选与鉴定[J].微生物学杂志,2013(5):18-24. |
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| 作者姓名： | 潘欣 蔡家麟 王颖 陈建霞 曾思良 方伟 陈敏 廖万清 余秀珍 张俊 宋维 |
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| 作者单位： | [1]上海俊维寓医馆,上海200433 [2]第二军医大学,上海200433 [3]上海交通大学基础医学院,上海200025 [4]上海市肺科医院,上海200433 |
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| 基金项目： | 国家重点基础研究发展计划（973）资助项目（2013CB531601）;国家自然科学基金（30972633） |
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| 摘 要： | 单域重链抗体是目前中和胞内病原体抗原的重要分子之一，研究以结核分枝杆菌Rv0733-6His融合抗原为靶标，对羊驼非免疫单域重链抗体库进行了3轮淘洗，通过ELISA和测序方法，从1024个克隆中筛选出10个独立单域重链抗体序列，继而用原核表达并鉴定了1株Rv0733-VHH-Fe-6His抗体。免疫印迹和免疫荧光结果均显示Rv0733-VHH—Fe-6His抗体可以特异性地结合Rv0733抗原。提示Rv0733-VHH抗体可能具备结合胞内菌相关抗原的潜力。
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| 关 键 词： | 结核分枝杆菌 表达纯化 单域重链抗体 噬菌体抗体库 |
Screening ＆ Identification of Mycobacterium tuberculosis Rv0733-6His Antigens from Alpaca Single-Domain Heavy-Chain Antibody |
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| PAN Xin,CAI Jia-lin,WANG Ying,CHEN Jian-xi,ZENG Si-liang,FANG Wei,CHEN Min,LIAO Wan-qing,YU Xiu-zhen,ZHANG Jun and SONG Wei.Screening ＆ Identification of Mycobacterium tuberculosis Rv0733-6His Antigens from Alpaca Single-Domain Heavy-Chain Antibody[J].Journal of Microbiology,2013(5):18-24. |
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| Authors: | PAN Xin CAI Jia-lin WANG Ying CHEN Jian-xi ZENG Si-liang FANG Wei CHEN Min LIAO Wan-qing YU Xiu-zhen ZHANG Jun and SONG Wei |
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| Institution: | 1 ( 1. Shanghai Junwei Fine Med. Club, Shanghai 200433 ; 2. 2nd Mil. Med. Uni. , Shanghai 200433 ; 3. Shanghai Jiao Tong Uni. ,Sch. of Med. , Shanghai 200025; 4. Shanghai Pulmonary Hosp. , Shanghai 200433 ) |
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| Abstract: | Single domain of heavy-chain antibody is one of the important molecules to neutralize intracellular pathogen antigens. In this study a blending antigen of Mycobacterium tuberculosis RvO733-6His was taken as a target to carry out three rounds of elutriation of alpaca non-immune single domain heavy chain antibody, and screened 10 independent single domain heavy-chain antibody sequence from 1 024 clones through ELISA and sequencing method, continued to identify a RvO733-VHH-Fc-6His antibody with prokaryotie expression. The results of both western blotting and immunofluorescence demonstrated that RvO733-VHH-Fc-6His antibody could specifically combine with Rv0733 antigen. Suggested that RvO733-VHH antibody may have a potential to bind intracellular pathogen related antigen. |
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| Keywords: | Mycobacterium tuberculosis expression purification single domain of heavy-chain antibody phage display antibody library |
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