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标题：Preparation of Specific Nanobodies and Their Application in the Rapid Detection of Nodularin-R in Water Samples
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作者：Jinyi Yang ; Rui Si ; Guangpei Wu 等
期刊名称：Foods
电子版ISSN：2304-8158
出版年度：2021
卷号：10
期号：11
DOI：10.3390/foods10112758
语种：English
出版社：MDPI Publishing
摘要：Nanobodies have several advantages, including great stability, sensibility, and ease of production; therefore, they have become important tools in immunoassays for chemical contaminants. In this manuscript, nanobodies for the detection of the toxin Nodularin-r (NOD-R), a secondary metabolite of cyanobacteria that could cause a safety risk for drinks and food for its strong hepatotoxicity, were for the first time selected from an immunized Bactrian camel VHH phage display library. Then, a sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for NOD-R, based on the nanobody N56 with great thermostability and organic solvent tolerance, was established under optimized conditions. The results showed that the limit of detection for NOD-R was 0.67 µg/L, and the average spike recovery rate was between 84.0 and 118.3%. Moreover, the ic-ELISA method was validated with spiked water sample and confirmed by UPLC–MS/MS, which indicated that the ic-ELISA established in this work is a reproducible detection assay for nodularin residues in water samples.
关键词：nodularin; nanobody; detection; ic-ELISA