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首页> 外文期刊>Chemistry: A European journal >Isolation, Purification, Characterization and Direct Conjugation of the Lipid A-Free Lipopolysaccharide of Vibrio cholerae O139
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Isolation, Purification, Characterization and Direct Conjugation of the Lipid A-Free Lipopolysaccharide of Vibrio cholerae O139

机译：血脂胆囊型钙脂多糖的分离，纯化，表征和直接共轭

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摘要

The lipopolysaccharide (LPS) of Vibrio cholerae O139, strain CIRS245, was isolated conventionally, and the lipidA was removed by mild acid hydrolysis (0.1m NaOAc buffer containing 1% SDS, pH 4.2, 95°C, 8 h). The crude product was a complex mixture consisting mainly of constituent fragments of the O-specific polysaccharide-core (OSPc). The OSPc was only a minor component in the mixture. Twostage purification of the crude OSPc by HPLC gave pure OSPc fragment of the LPS, as shown by NMR spectroscopy, analytical HPLC and ESI-MS. This material is the purest OSPc fragment of the LPS from Vibrio cholerae O139 reported to date. The purified OSPc was readily converted to the corresponding methyl squarate derivative and the latter was conjugated to BSA. The conjugate, when examined by ELISA, showed immunoreactivity with sera from patients in Bangladesh recovering from cholera caused by V. cholerae O139, but not O1.
机译：常规分离vibrio霍乱O139，菌株CiRS245的脂多糖（LPS），并通过温和的酸水解除去脂质（0.1M Naoac缓冲液，含有1％SDS，pH 4.2,95℃，8h）。 粗产物是一种复杂的混合物，主要是O特异性多糖 - 核（OSPC）的组成片段组成。 OSPC仅是混合物中的次要组件。 通过HPLC纯化粗OSPC的纯化给出了LPS的纯OSPC片段，如NMR光谱，分析HPLC和ESI-MS所示。 该材料是来自迄今为止的富轴霍乱O139的LPS最纯的OSPC片段。 将纯化的OSPC容易地转化为相应的甲基平方衍生物，后者与BSA缀合。 当ELISA检查时，缀合物显示出免疫反应与来自孟加拉国患者的血清从霍乱患者中恢复，由V.Cholerae O139引起的，但不是O1。

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  • 来源
    《Chemistry: A European journal》 |2019年第56期|共11页
  • 作者

    Peng Xu; Jana Korcová; Peter Baráth; Al?beta ?í?ová; Jana Valáriková; Firdausi Qadri; Meagan Kelly; Robert D. OConnor; Edward T. Ryan; Slavomír Bystricky; Pavol Ková?;

  • 作者单位

    Laboratory of Bioorganic Chemistry National Institute of Diabetes and Digestive and Kidney Diseases National Institutes of Health 9000 Rockville Bethesda MD 20892-0815 (USA);

    Institute of Chemistry Slovak Academy of Sciences 84238 Bratislava (Slovak Republic);

    Institute of Chemistry Slovak Academy of Sciences 84238 Bratislava (Slovak Republic);

    Institute of Chemistry Slovak Academy of Sciences 84238 Bratislava (Slovak Republic);

    Institute of Chemistry Slovak Academy of Sciences 84238 Bratislava (Slovak Republic);

    Infectious Diseases Division International Centre for Diarrhoeal Disease Research Bangladesh (icddr b) Dhaka (Bangladesh);

    Division of Infectious Diseases Massachusetts General Hospital Boston MA 02114 (USA);

    Laboratory of Bioorganic Chemistry National Institute of Diabetes and Digestive and Kidney Diseases National Institutes of Health 9000 Rockville Bethesda MD 20892-0815 (USA);

    Division of Infectious Diseases Massachusetts General Hospital Boston MA 02114 (USA);

    Institute of Chemistry Slovak Academy of Sciences 84238 Bratislava (Slovak Republic);

    Laboratory of Bioorganic Chemistry National Institute of Diabetes and Digestive and Kidney Diseases National Institutes of Health 9000 Rockville Bethesda MD 20892-0815 (USA);

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 应用化学;
  • 关键词

    glycoconjugate vaccines; NMR spectroscopy; polysaccharides; squaric acid; Vibrio cholerae;

    机译：糖醌疫苗;NMR光谱;多糖;种族酸;霍乱霍乱;

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