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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 heavy chain antibody is currently one of the important molecules in neutralizing intracellular pathogen antigens. In this study, three cycles of panning of the non-immunized single-chain heavy chain antibody library of alpaca were performed with Mycobacterium tuberculosis Rv0733-6His fusion antigen as target , 10 independent single domain heavy chain antibody sequences were screened from 1024 clones by ELISA and sequencing, and then one Rv0733-VHH-Fc-6His antibody was expressed and identified by prokaryotic expression. Immunoblotting and immunofluorescence results showed that the Rv0733-VHH-Fc-6His antibody specifically binds to the Rv0733 antigen. Suggesting that the Rv0733-VHH antibody may have the potential to bind to intracellular bacteria-associated antigens.