抗金刚烷胺和金刚乙胺单克隆抗体的制备及用于检测鸡肉和肝脏中金刚烷胺和金刚乙胺的间接竞争酶联免疫吸附测定法的建立
Preparation of a monoclonal antibody against amantadine and rimantadine and development of an indirect competitive enzyme-linked immunosorbent assay for detecting the same in chicken muscle and liver.
作者信息
Peng Dapeng, Wei Wei, Pan Yuanhu, Wang Yulian, Chen Dongmei, Liu Zhenli, Wang Xu, Dai Menghong, Yuan Zonghui
机构信息
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, MOA Laboratory for Risk Assessment of Quality and Safety of Livestock and Poultry Products, Hubei Collaborative Innovation Centre for Animal Nutrition and Feed Safety, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, MOA Laboratory for Risk Assessment of Quality and Safety of Livestock and Poultry Products, Hubei Collaborative Innovation Centre for Animal Nutrition and Feed Safety, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
出版信息
J Pharm Biomed Anal. 2017 Jan 30;133:56-63. doi: 10.1016/j.jpba.2016.11.009. Epub 2016 Nov 10.
A monoclonal antibody (mAb) was produced in order to monitor the illegal use of amantadine and rimantadine in animals. The produced mAb 2G3 exhibited an IC value of 15.8μgL for amantadine and exhibited cross-reactivity to both amantadine (100%) and rimantadine (70.6%). Standard curves ranged from 5 to 80μgL for 2G3. The limits of detection of the developed indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) ranged from 5.0μgkg to 5.4μgkg in chicken muscle and liver. The recoveries were 81.3% to 98.1% with a coefficient of variation less than 15.7%. Good correlations were observed between the results of the ic-ELISA and liquid chromatography-mass spectrometry in the incurred tissues. These results suggest that ic-ELISA is a sensitive, accurate, and low-cost method that could be a useful tool for screening the residues of amantadine and rimantadine in chicken muscle and liver.
为监测金刚烷胺和金刚乙胺在动物中的非法使用，制备了一种单克隆抗体（mAb）。所制备的单克隆抗体2G3对金刚烷胺的半数抑制浓度（IC值）为15.8μg/L，对金刚烷胺（100%）和金刚乙胺（70.6%）均表现出交叉反应性。2G3的标准曲线范围为5至80μg/L。所建立的间接竞争酶联免疫吸附测定法（ic-ELISA）在鸡肌肉和肝脏中的检测限为5.0μg/kg至5.4μg/kg。回收率为81.3%至98.1%，变异系数小于15.7%。在实际受检组织中，ic-ELISA结果与液相色谱-质谱法结果之间观察到良好的相关性。这些结果表明，ic-ELISA是一种灵敏、准确且低成本的方法，可作为筛查鸡肌肉和肝脏中金刚烷胺和金刚乙胺残留的有用工具。
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