State Key Laboratory of Food Science and Technology, and Sino-German Joint Research Institute, Nanchang University, 235 Nanjing East Road, Nanchang 330047, China.
State Key Laboratory of Food Science and Technology, and Sino-German Joint Research Institute, Nanchang University, 235 Nanjing East Road, Nanchang 330047, China.
Food Chem. 2020 Jul 30;319:126546. doi: 10.1016/j.foodchem.2020.126546. Epub 2020 Mar 2.
A rapid and sensitive immunochromatographic strip (ICS) based on a single-chain variable fragment (scFv) was developed for detecting fumonisin B (FB). The ICS was based on a competitive reaction for colloidal gold-labeled scFv between FB and FB-BSA, which was used along with sheep anti-mouse IgG as capture reagents immobilized at test and control lines, respectively, on a nitrocellulose membrane of the strip. The limit of detection of the ICS was 2.5 ng/mL (25 μg/kg) FB in buffer, and the sensitivity was eight times higher than that of monoclonal antibodies for the preparation of the scFv. The cross-reactivity of the scFv with common mycotoxins was determined by ICS, the results showed that the scFv were not against other mycotoxins. Eight naturally contaminated maize samples were analyzed with the scFv-based ICS and by LC-MS/MS. The results of analysis obtained with the strip assay showed good agreement with those obtained by LC-MS/MS.
基于单链可变片段 (scFv) 的快速灵敏免疫层析条（ICS）已被开发用于检测伏马菌素 B（FB）。ICS 基于胶体金标记的 scFv 与 FB 和 FB-BSA 之间的竞争反应，该反应与 FB-BSA 一起使用，作为捕获试剂，分别固定在测试线和对照线上，位于条带的硝酸纤维素膜上。ICS 的检测限为 2.5ng/mL（25μg/kg）缓冲液中的 FB，其灵敏度比用于制备 scFv 的单克隆抗体高 8 倍。通过 ICS 确定了 scFv 与常见霉菌毒素的交叉反应性，结果表明 scFv 对其他霉菌毒素没有反应。用基于 scFv 的 ICS 和 LC-MS/MS 分析了 8 个受自然污染的玉米样品。条带分析获得的结果与 LC-MS/MS 获得的结果吻合良好。
