Preparation of a monoclonal antibody and development of an indirect competitive ELISA for the detection of chlorpromazine residue in chicken and swine liver.
Preparation of a monoclonal antibody and development of an indirect competitive ELISA for the detection of chlorpromazine residue in chicken and swine liver.
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DOI：
10.1002/jsfa.4014
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发表时间：
2010-08
影响因子：
4.1
通讯作者：
W. Liu;Weihua Li;Weiwei Yin;Meng Meng-Meng;Yuping Wan;Caiwei Feng;Shanliang Wang;Rimo Xi
中科院分区：
文献类型：
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作者：
W. Liu;Weihua Li;Weiwei Yin;Meng Meng-Meng;Yuping Wan;Caiwei Feng;Shanliang Wang;Rimo Xi
背景氯丙嗪是一种典型的抗精神病药物，作为饲料添加剂用于使食用动物镇静和促进生长。氯丙嗪在动物体内积聚会对循环系统和神经系统造成副作用，并对血细胞、皮肤和眼睛造成不良影响。为检测食品动物中氯丙嗪残留，在制备抗氯丙嗪单克隆抗体的基础上，建立了间接竞争酶联免疫吸附试验（ELISA）。结果以阳离子牛血清白蛋白（cBSA）为免疫原，偶联氯丙嗪制备的抗体对氯丙嗪具有较高的敏感性，IC（50）值为0.73ppb。将该方法应用于猪肝和鸡肉样品中氯丙嗪的检测，结果满意。在鸡肝和猪肝中的回收率分别为88-95%和86- 95%，批内变异系数分别< 15.3%和<13.5%。结论建立了一种灵敏度高、特异性强的抗氯丙嗪单克隆抗体间接竞争ELISA检测方法。本研究提供的免疫分析方法有望取代色谱光谱法用于食品动物中氯丙嗪残留的监管分析。
BACKGROUND Chlorpromazine is a typical antipsychotic drug used to make food-producing animals calm and promote growth as feed additives. Accumulation of chlorpromazine in animal bodies would cause side effects in the circulatory and nervous systems, and have adverse effects on blood cells, the skin and the eye. To detect the chlorpromazine residue in food producing animals, an indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed based on preparation of an anti-chlorpromazine monoclonal antibody. RESULTS The antibody generated from immunogen of cationic bovine serum albumin (cBSA) coupled with chlorpromazine showed high sensitivity toward chlorpromazine with an IC(50) value of 0.73 ppb. The ELISA method was applied to detect swine liver and chicken samples spiked by chlorpromazine and satisfactory results were obtained. The recovery rates in chicken and swine liver were in the range of 88-95% and 86-95%, respectively; the intra-assay coefficients of variation were both < 15.3% and < 13.5%, respectively. CONCLUSION An indirect competitive ELISA method based on a monoclonal antibody towards chlorpromazine with excellent sensitivity and specificity has been successfully developed. The immunoassay provided in this study was a hopeful alternative to chromatography spectrometry for regulatory analysis of chlorpromazine residue in food-producing animals.