Vibrio cholerae, causing the life-threatening diarrheal disease cholerae, can be divided into different serogroups based on the structure of lipopolysaccharide (LPS) that consists of lipid-A, core-polysaccharide and O-antigen polysaccharide (O-PS). The O1 serogroup, the predominant cause of cholera, includes two major serotypes, Inaba and Ogawa. These serotypes are differentiated by the presence of a single 2-O-methyl group in the upstream terminal perosamine of the Ogawa O-PS, which is absent in the Inaba O-PS. To ensure the consistent quality and efficacy of the current cholera vaccines, accurate measurement and characterization of each of these two serotypes is highly important. In this study, we efficiently screened a phage-displayed human synthetic Fab library by bio-panning against Ogawa LPS and finally selected three unique mAbs (D9, E11 and F7) that specifically reacts with Ogawa LPS. The mAbs bound to Vibrio cholerae vaccine in a dose-dependent fashion. Sequence and structure analyses of antibody paratopes suggest that IgG D9 might have the same fine specificity as that of the murine mAbs, which were shown to bind to the upstream terminal perosamine of Ogawa O-PS, whereas IgGs F7 and E11 showed some different characteristics in the paratopes. To our knowledge, this study is the first to show the generation of Ogawa-specific mAbs using phage display technology. The mAbs will be useful for identification and quantification of Ogawa LPS in multivalent V. Cholerae vaccines.

中文翻译：

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从噬菌体展示的人类合成 Fab 文库中产生和表征霍乱弧菌 O1 的 Ogawa 脂多糖单克隆抗体。

霍乱弧菌可引起危及生命的腹泻病霍乱，根据脂多糖（LPS）的结构可分为不同的血清群，脂多糖由脂质 A、核心多糖和 O 抗原多糖（O-PS）组成。O1 血清群是霍乱的主要原因，包括两种主要血清型，Inaba 和 Ogawa。这些血清型通过存在单个 2- O-Ogawa O-PS 上游末端过氧化胺中的甲基，Inaba O-PS 中不存在。为确保当前霍乱疫苗的质量和功效始终如一，准确测量和表征这两种血清型非常重要。在这项研究中，我们通过针对 Ogawa LPS 的生物淘选有效筛选了噬菌体展示的人类合成 Fab 文库，并最终选择了三种与 Ogawa LPS 特异性反应的独特 mAb（D9、E11 和 F7）。与霍乱弧菌结合的 mAb疫苗以剂量依赖的方式。抗体互补位的序列和结构分析表明，IgG D9 可能具有与小鼠单克隆抗体相同的优良特异性，后者显示与 Ogawa O-PS 的上游末端过氧化胺结合，而 IgG F7 和 E11 显示出一些不同的特征互补位。据我们所知，这项研究首次展示了使用噬菌体展示技术生成 Ogawa 特异性 mAb。mAb 可用于鉴定和定量多价霍乱弧菌疫苗中的 Ogawa LPS。