中文摘要：模拟表位，或表位模拟，可作为毒素结合物的替代品，用于竞争性免疫分析，检测低分子天然毒物。本研究开发了一种用噬菌体展示技术筛选的新型模拟表位检测伏马菌素B1的芯片免疫分析法。研究表明该方法能在欧盟法规设定的水平上量化霉菌毒素，并有望适应将来包括其他霉菌毒素的多重检测。
外文摘要：Mimotopes, or epitope mimics, can be applied to competitive immunoassays, for the detection of low molecular weight natural toxicants, as an alternative to toxin-conjugates. In this work, we report the development of a microarray-based immunoassay for the detection of fumonisin B-1 using a novel mimotope selected by phage display technology. Fumonisin-specific antibody was used to isolate mimotopes from a 12-mer peptide library in successive selection rounds. Enrichment of antibody binding phages was observed after three panning rounds, and sequence analysis of randomly selected monoclonal phages revealed two conserved peptide sequences. Clone A2, with peptide sequence VTPNDDTFDPFR, showed the best response in enzyme-linked immunosorbent assay (ELISA) in terms of sensitivity and reproducibility and was selected for microarray development. A biotinylated synthetic derivative of this mimotope was immobilized onto epoxy-glass slides, and fumonisin B-1 was detected in a competitive binding inhibition assay using the antifumonisin antibody and a labeled secondary antibody. The array showed an IC50 value of 37.1 +/- 2.4 ng (n = 9), a detection limit of 11.1 ng mL(-1) and a dynamic range from 17.3 to 79.6 ng mL(-1). Good specificity toward fumonisin B-1 and its structural analog, fumonisin B-2, was observed, together with negligible cross-reactivity for other mycotoxins produced by the same fungi species. The,mimotope microarray was applied to the analysis of fumonisin B-1. in spiked maize and wheat samples. The method enabled quantification of the mycotoxin at the levels set by European legislation and holds promise for future adaptation to include other mycotoxins for multiplex detection.
作者：Peltomaa, R；Benitoyelia-Pena, E；Barderas, R；Sauer, U；Andrade, MG；Moreno-Bondi, MC
作者单位：Univ Complutense Madrid
期刊名称：ANALYTICAL CHEMISTRY
期刊影响因子：5.886
出版年份：2017
出版刊次：11
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