Monoclonal antibody-based indirect competitive ELISA for quantitative detection of Enterobacteriaceae siderophore enterobactin

文献类型： 外文期刊

第一作者： Cui, Yifang

作者： Cui, Yifang;Guo, Fangfang;Cao, Xiaoya;Wang, Xue;Xu, Fuzhou;Wang, Huiwen;Zeng, Ximin;Lin, Jun;Cui, Guolin

作者机构：

关键词： Enterobacteriaceae; Enterobactin; Monoclonal antibody; ic-ELISA

期刊名称：FOOD CHEMISTRY （ 影响因子：9.231； 五年影响因子：8.795 ）

ISSN： 0308-8146

年卷期： 2022 年 391 卷

页码：

收录情况： SCI

摘要： Enterobactin (Ent) is a promising indicator to monitor intestinal level of Enterobacteriaceae for assessment of gut inflammation. In this study, we developed a monoclonal antibody (mAb)-based ELISA for Ent quantification. We immunized mice with an Ent conjugate vaccine. An mAb named 2E4, with the highest anti-Ent antibody titer, was selected for developing indirect competitive ELISA (ic-ELISA). The purified mAb 2E4 showed high affinity (3.1 x 10(-10) M) and specificity to Ent. The limit of detection of ic-ELISA was 0.39 mu g/mL. The intra-and inter assay recovery rates of standard curve were up to 94.6% with the coefficients of variation between 4.0% and 12.3%, indicating high accuracy, repeatability, and reproducibility of the ic-ELISA. In addition, the ic-ELISA was able to quantitatively detect Ent produced in different bacterial cultures. Collectively, this study developed an icELISA with excellent performance in Ent quantification, laying a solid foundation for Ent-based diagnostics of gut health.

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