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结核分枝杆菌Rv0733-6His抗原羊驼单域重链抗体的筛选与鉴定

Screening & Identification of Mycobacterium tuberculosis Rv0733-6His Antigens from Alpaca Single-Domain Heavy-Chain Antibody
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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1 024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-Fc-6His抗体。免疫印迹和免疫荧光结果均显示Rv0733-VHH-Fc-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。 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.
作者 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
机构地区 上海俊维寓医馆 第二军医大学 上海交通大学基础医学院 上海市肺科医院
出处 《微生物学杂志》 CAS CSCD 2013年第5期18-24,共7页 Journal of Microbiology
基金 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)
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 Mycobacterium tuberculosis expression purification single domain of heavy-chain antibody phage display antibody library
分类号 R378.911 [医药卫生—病原生物学]
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参考文献9

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二级参考文献25

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微生物学杂志
2013年 第5期

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