College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, Hubei, China.
Institute of Quality Standard and Testing Technology for Agro-Products, Hubei Academy of Agricultural Sciences, Wuhan, 430064, Hubei, China.
Anal Bioanal Chem. 2019 Jul;411(17):3881-3890. doi: 10.1007/s00216-019-01862-7. Epub 2019 May 31.
Aflatoxin B (AFB) is one of the major mycotoxins, which naturally occurs in food and agricultural products. In this study, a cyclic peptide (CVPSKPGLC) mimicking AFB was used to develop a biotinylated peptide-based immunoassay (bp-ELISA) for AFB determination. This cyclic peptide was isolated from a commercially available phage-displayed random 7-mer cyclic peptide library, and then synthesized chemically. Instead of phage particles, the peptide was biotinylated and used to detect AFB by bp-ELISA, with an IC of 0.92 ng/mL, which was approximately 60-fold better than that of phage ELISA. Good recoveries (83-102%) were obtained in spiked rice and corn samples, which were further validated by high-performance liquid chromatography-fluorescence detector. As better sensitivities (0.92-1.21 ng/mL) were obtained by bp-ELISA even using selected anti-AFB antibodies prepared previously in laboratory, this cyclic peptide is suitable as a substitute for synthetic competitive AFB antigens in food contamination monitoring. Graphical abstract.
黄曲霉毒素 B（AFB）是一种主要的霉菌毒素，天然存在于食品和农产品中。在本研究中，一种模拟 AFB 的环肽（CVPSKPGLC）被用于开发用于检测 AFB 的生物素化肽基免疫测定法（bp-ELISA）。该环肽是从市售的噬菌体展示的随机 7 -mer 环肽文库中分离出来的，然后通过化学合成得到。该肽被生物素化而不是噬菌体颗粒，并用于通过 bp-ELISA 检测 AFB，其 IC50 为 0.92ng/mL，比噬菌体 ELISA 约高 60 倍。在添加的大米和玉米样品中获得了良好的回收率（83-102%），并通过高效液相色谱-荧光检测器进一步验证。由于使用之前在实验室中制备的选定抗 AFB 抗体，bp-ELISA 甚至获得了更好的灵敏度（0.92-1.21ng/mL），因此这种环肽适合用作食品污染监测中合成竞争 AFB 抗原的替代品。
