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