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用于快速检测萘普生的半抗原设计、单克隆抗体制备及免疫分析方法开发

Hapten design, monoclonal antibody preparation, and immunoassay development for rapid detection of naproxen.

作者信息

Liu Qingqing, Xu Xinxin, Wu Aihong, Xu Chuanlai, Kuang Hua

机构信息

State Key Lab of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, People's Republic of China.

出版信息

J Mater Chem B. 2025 May 7;13(18):5417-5426. doi: 10.1039/d5tb00330j.

Abstract

Naproxen (NPX) is a non-steroidal anti-inflammatory drug that poses a significant environmental threat due to widespread use and resistance to degradation. Here, a high-affinity monoclonal antibody (mAb) against NPX was prepared by computer-aided hapten design, mouse immunization, and cell fusion. The conformation of the hapten predicted by computer-assisted molecular simulation was consistent with the mouse immunization results. A highly sensitive immunochromatographic assay (ICA) was established based on this mAb for the detection of NPX in sewage, soil and serum with cut-off values of 0.5 ng mL, 2 μg kg, and 5 ng mL, detection limits of 0.010 ng mL, 0.051 μg kg, and 0.028 ng mL, and recovery rates of 90.3-98.1%, 90.3-106.5%, and 96.9-106.0%, respectively. The detection results of real samples were highly consistent with LC-MS/MS, thereby demonstrating reliability of the proposed ICA.

摘要

萘普生(NPX)是一种非甾体抗炎药,因其广泛使用和抗降解性而对环境构成重大威胁。在此,通过计算机辅助半抗原设计、小鼠免疫和细胞融合制备了一种针对NPX的高亲和力单克隆抗体(mAb)。计算机辅助分子模拟预测的半抗原构象与小鼠免疫结果一致。基于该mAb建立了一种高灵敏度免疫色谱法(ICA),用于检测污水、土壤和血清中的NPX,其截断值分别为0.5 ng/mL、2 μg/kg和5 ng/mL,检测限分别为0.010 ng/mL、0.051 μg/kg和0.028 ng/mL,回收率分别为90.3 - 98.1%、90.3 - 106.5%和96.9 - 106.0%。实际样品的检测结果与液相色谱 - 串联质谱法(LC - MS/MS)高度一致,从而证明了所提出的ICA的可靠性。

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