筛选特异性纳米抗体以开发检测牛奶中金黄色葡萄球菌的免疫分析方法。
Selection of specific nanobodies to develop an immuno-assay detecting Staphylococcus aureus in milk.
机构信息
Tianjin Key Laboratory of Food Science and Health, School of Medicine, Nankai University, Tianjin 300071, China.
College of Food Science and Engineering, Tianjin University of Science & Technology, Tianjin 300457, China.
出版信息
Food Chem. 2021 Aug 15;353:129481. doi: 10.1016/j.foodchem.2021.129481. Epub 2021 Mar 6.
The interaction between conventional immunoglobulins (Igs) and the Ig-binding surface proteins of Staphylococcus aureus (S. aureus) have obstructed the development of immuno-assays to detect these bacteria. The current study aimed to select nanobodies (Nbs) recognizing specifically S. aureus and to establish an immuno-assay to uncover S. aureus contaminations in foods. An alpaca was immunized with an inactivated S. aureus strain followed by the construction of a Nb library from which four target-specific Nbs were retrieved. Subsequently, a sandwich ELISA employing the Nb147 and biotinylated-Nb147 pair to capture and to detect S. aureus, respectively, was established to possess a detection limit of 1.4 × 10 colony forming units (CFU)/mL. The dedicated immuno-assay has been verified by detecting 10 CFU/mL of S. aureus in milk samples after an 8 h-enrichment step. This study provides the basis of an easy, reproducible and effective immuno-assay to screen for S. aureus contaminations in foods.
常规免疫球蛋白（Igs）与金黄色葡萄球菌（S. aureus）的 Ig 结合表面蛋白之间的相互作用，阻碍了免疫分析检测这些细菌的发展。本研究旨在筛选特异性识别金黄色葡萄球菌的纳米抗体（Nbs），并建立一种免疫分析方法来检测食品中的金黄色葡萄球菌污染。用灭活的金黄色葡萄球菌菌株对一只羊驼进行免疫，然后从该菌株构建一个纳米抗体文库，从中回收了 4 种靶向特异性纳米抗体。随后，建立了一种夹心 ELISA 方法，使用纳米抗体 147 和生物素化纳米抗体 147 分别用于捕获和检测金黄色葡萄球菌，其检测限为 1.4×10 个菌落形成单位（CFU）/mL。该专用免疫分析方法已通过在 8 小时富集步骤后在牛奶样本中检测到 10 CFU/mL 的金黄色葡萄球菌得到验证。本研究为在食品中筛选金黄色葡萄球菌污染提供了一种简单、可重复和有效的免疫分析方法的基础。
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