State Key Laboratory of Magnetic Resonance and Atomic Molecular Physics, Key Laboratory of Magnetic Resonance in Biological Systems, National Center for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences-Wuhan National Laboratory for Optoelectronics, Wuhan 430071, China.
Department of Chemistry, College of Science, Huazhong Agricultural University, Wuhan 430070, China.
Biosensors (Basel). 2023 May 30;13(6):593. doi: 10.3390/bios13060593.
Forchlorfenuron (CPPU) is a widely used plant growth regulator in agriculture, and CPPU residue in food can cause harm to human health. Thus, it is necessary to develop a rapid and sensitive detection method for CPPU monitoring. In this study, a new monoclonal antibody (mAb) against CPPU with high affinity was prepared by a hybridoma technique, and a magnetic bead (MB)-based analytical method was established for the determination of CPPU by a one-step procedure. Under optimized conditions, the detection limit of the MB-based immunoassay was as low as 0.0004 ng/mL, which was five times more sensitive than the traditional indirect competitive ELISA (icELISA). In addition, the detection procedure took less than 35 min, a significant improvement over the 135 min required for icELISA. The selectivity test of the MB-based assay also showed negligible cross-reactivity with five analogues. Furthermore, the accuracy of the developed assay was assessed by the analysis of spiked samples, and the results agreed well with those obtained by HPLC. The excellent analytical performance of the proposed assay suggests its great potential for routine screening of CPPU, and it provides a basis for promoting the application of more immunosensors in the quantitative detection of low concentrations of small organic molecules in food.
氯吡脲（CPPU）是农业中广泛使用的植物生长调节剂，食品中的 CPPU 残留会对人体健康造成危害。因此，有必要开发一种快速灵敏的 CPPU 监测方法。本研究采用杂交瘤技术制备了一种对 CPPU 具有高亲和力的单克隆抗体（mAb），并建立了一种基于磁性珠（MB）的分析方法，通过一步法测定 CPPU。在优化条件下，MB 免疫分析的检测限低至 0.0004ng/mL，比传统的间接竞争 ELISA（icELISA）灵敏 5 倍。此外，检测过程耗时不到 35 分钟，比 icELISA 所需的 135 分钟有显著的改善。MB 分析方法的选择性测试也显示与五种类似物的交叉反应可以忽略不计。此外，通过对加标样品的分析评估了所开发分析方法的准确性，结果与 HPLC 法的结果吻合良好。该分析方法具有出色的分析性能，表明其在 CPPU 的常规筛选中具有很大的潜力，为促进更多免疫传感器在食品中低浓度小分子的定量检测中的应用提供了依据。
