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基于抗(-)-沙丁胺醇手性单克隆抗体的超灵敏磁辅助侧向流动免疫分析方法,用于检测猪尿液和猪肉中的 38 种β-激动剂

Ultrasensitive Magnetic Assisted Lateral Flow Immunoassay Based on Chiral Monoclonal Antibody against -(-)-Salbutamol of Broad-Specificity for 38 β-Agonists Detection in Swine Urine and Pork.

机构信息

Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University, Guangzhou 510642, China.

Wuzhou Institute for Food and Drug Control, Wuzhou 543099, China.

出版信息

J Agric Food Chem. 2022 Apr 6;70(13):4112-4122. doi: 10.1021/acs.jafc.2c00875. Epub 2022 Mar 28.

Abstract

Screening for ″zero tolerance″ β-agonists requires broad-specificity and sensitivity methods. Herein, -(-)-salbutamol (SAL) is chirally separated and designed as a hapten, and a monoclonal antibody (mAb) was first prepared with an IC of 0.27 ng/mL, which can recognize 38 β-agonists simultaneously. The broad-specificity of chiral mAb was explored by molecular simulation technology. Magnetic nanoparticles (MNPs) were then synthesized and applied as a signal tracer to develop a lateral flow immunoassay (LFIA). The limits of detection of MNPs-LFIA for SAL in swine urine and pork were 0.05 and 0.09 μg/kg, which was (2-125)-fold lower than that of the reported LFIAs. The recoveries were between 95.8 and 116.7%, with the coefficient of variation from 2.7 to 15.4%. Parallel analysis of 44 samples by commercial ELISA kits confirmed the reliability. Therefore, our work not only provides a broad-specificity and ultrasensitive method for β-agonists but also suggests that chirality is the new general theory that guided the rational hapten design.

摘要

筛查“零容忍”β-激动剂需要广谱特异性和灵敏度方法。本文中,(-)-沙丁胺醇(SAL)被手性拆分并设计为半抗原,首次制备出 IC₅₀为 0.27ng/mL 的单克隆抗体(mAb),可同时识别 38 种β-激动剂。通过分子模拟技术探索了手性 mAb 的广谱特异性。然后合成磁性纳米粒子(MNPs)并用作信号示踪剂,以开发侧向流动免疫分析(LFIA)。MNPs-LFIA 检测猪尿和猪肉中 SAL 的检出限分别为 0.05 和 0.09μg/kg,比报道的 LFIA 低(2-125)倍。回收率在 95.8%至 116.7%之间,变异系数为 2.7%至 15.4%。通过商业 ELISA 试剂盒对 44 个样本进行平行分析证实了该方法的可靠性。因此,我们的工作不仅为β-激动剂提供了一种广谱特异性和超灵敏的方法,还表明手性是指导合理半抗原设计的新通用理论。

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