Cheng Guyue, Li Feng, Peng Dapeng, Huang Lingli, Hao Haihong, Liu Zhenli, Wang Yulian, Yuan Zonghui
MOA Laboratory for Risk Assessment of Quality and Safety of Livestock and Poultry Products, Huazhong Agricultural University, Wuhan 430070, China.
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for Detection of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan 430070, China.
Anal Biochem. 2014 Aug 15;459:18-23. doi: 10.1016/j.ab.2014.05.005. Epub 2014 May 20.
β-Adrenergic agonists (β-agonists) are illegally used in animal husbandry, threatening the health of consumers. To realize multianalyte detection of β-agonists, a β2-adrenergic receptor (β2-AR) was cloned from Syrian hamster lung and heterogeneously expressed by Spodoptera frugiperda (Sf9) cells. The recombinant β2-AR was purified from intracellular soluble proteins of infected Sf9 cells, and was utilized to establish an enzyme-linked-receptor assay (ELRA) to detect a group of β-agonists simultaneously. This assay was based on direct competitive inhibition of binding of horseradish peroxidase-labeled ractopamine to the immobilized β2-AR proteins by β-agonists. The IC50 and limit of detection values for ractopamine were 30.38μgL(-1) and 5.20μgL(-1), respectively. Clenbuterol and salbutamol showed 87.7% and 58.5% cross-reactivities with ractopamine, respectively. This assay is simple, rapid, and environmentally friendly, showing a potential application in the screening of β-agonists in animal feeds.
β-肾上腺素能激动剂(β-激动剂)被非法用于畜牧业,威胁消费者健康。为实现β-激动剂的多分析物检测,从叙利亚仓鼠肺中克隆了β2-肾上腺素能受体(β2-AR),并由草地贪夜蛾(Sf9)细胞进行异源表达。重组β2-AR从感染的Sf9细胞的细胞内可溶性蛋白中纯化出来,并用于建立酶联受体分析(ELRA)以同时检测一组β-激动剂。该分析基于β-激动剂对辣根过氧化物酶标记的莱克多巴胺与固定化β2-AR蛋白结合的直接竞争性抑制。莱克多巴胺的IC50和检测限分别为30.38μg L(-1)和5.20μg L(-1)。克伦特罗和沙丁胺醇与莱克多巴胺的交叉反应率分别为87.7%和58.5%。该分析方法简单、快速且环保,在动物饲料中β-激动剂的筛选方面具有潜在应用价值。