School of Food Science and Engineering, Hainan University, Haikou 570228, China.
School of Food Science and Engineering, Hainan University, Haikou 570228, China.
Food Chem. 2024 Jun 15;443:138569. doi: 10.1016/j.foodchem.2024.138569. Epub 2024 Jan 29.
Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin and seriously threatens food safety, which requires rapid and sensitive detection methods for monitoring ZEN in agro-products. Herein, an alkaline phosphatase-tagged single-chain variable fragment fusion protein (ALP-scFv) was used as a bifunctional tracer to develop a colorimetric enzyme immunoassay (CEIA) and a chemiluminescent enzyme immunoassay (CLEIA) for ZEN. In addition, the interactions between scFv and ZEN were exploited by computer-assisted simulation, and four key amino acid sites were preliminarily identified. After optimization, the CEIA and CLEIA exhibited a limit of detection of 0.02 and 0.006 ng/mL, respectively. Furthermore, both methods showed favorable accuracy in recovery experiments and good selectivity in cross reactions. Moreover, the detection results of the actual samples from both methods correlated well with those from high-performance liquid chromatography. Overall, the ALP-scFv fusion tracer-based CEIA and CLEIA are demonstrated as reliable tools for ZEN detection in food.
玉米赤霉烯酮（ZEN）是一种非甾体类雌激素真菌毒素，严重威胁食品安全，因此需要快速灵敏的检测方法来监测农产品中的 ZEN。本研究中，我们将碱性磷酸酶标记的单链可变片段融合蛋白（ALP-scFv）用作双功能示踪剂，建立了用于 ZEN 的比色酶免疫分析（CEIA）和化学发光酶免疫分析（CLEIA）。此外，通过计算机辅助模拟研究了 scFv 与 ZEN 的相互作用，并初步鉴定了四个关键的氨基酸位点。经过优化，CEIA 和 CLEIA 的检测限分别为 0.02 和 0.006 ng/mL。此外，两种方法在回收率实验中均表现出良好的准确性，并且在交叉反应中具有良好的选择性。此外，两种方法对实际样品的检测结果与高效液相色谱法的检测结果相关性良好。总之，基于 ALP-scFv 融合示踪剂的 CEIA 和 CLEIA 可作为食品中 ZEN 检测的可靠工具。
