抗氯硝西泮单克隆抗体的制备及其在猪组织中苯二氮䓬类药物免疫分析中的应用
Production of monoclonal antibody against clonazepam for immunoassay of benzodiazepine drugs in swine tissues.
作者信息
Shan Wen C, Cui Ya L, He Xin, Zhang Lei, Liu Jing, Wang Jian P
机构信息
a College of Veterinary Medicine , Agricultural University of Hebei , Baoding , Hebei , China.
出版信息
J Environ Sci Health B. 2015;50(1):15-22. doi: 10.1080/03601234.2015.964620.
The objective of the present study was to produce a generic monoclonal antibody for immunoassay of residues of benzodiazepine drugs in swine tissues. Clonazepam was used to synthesize a hapten that was coupled to bovine serum albumin as an immunogen for the production of monoclonal antibody. Results showed that the obtained monoclonal antibody was able to recognize five benzodiazepine drugs simultaneously (clonazepam, flunitrazepam nitrazepam, diazepam, and oxazepam). The cross-reactivities were in the range of 24-100% and the limits of detection were in the range of 0.2-1.5 ng mL(-1) depending on the drug. Then a competitive indirect enzyme-linked immunosorbent assay was developed to determine the residues of five benzodiazepines in swine tissues (muscle, liver and kidney). The recoveries of five analytes from the fortified blank samples were in the range of 74.5-96.5% with coefficients of variation lower than 16.7%. Therefore, this immunoassay could be used as a rapid and simple method for the screening of residues of five benzodiazepine drugs in animal-derived foods.
本研究的目的是制备一种通用单克隆抗体，用于免疫测定猪组织中苯二氮䓬类药物的残留量。使用氯硝西泮合成半抗原，并将其与牛血清白蛋白偶联作为免疫原，用于制备单克隆抗体。结果表明，所获得的单克隆抗体能够同时识别五种苯二氮䓬类药物（氯硝西泮、氟硝西泮、硝西泮、地西泮和奥沙西泮）。交叉反应率在24%至100%之间，检测限在0.2至1.5 ng mL(-1)之间，具体取决于药物种类。然后开发了一种竞争性间接酶联免疫吸附测定法，用于测定猪组织（肌肉、肝脏和肾脏）中五种苯二氮䓬类药物的残留量。从加标空白样品中回收的五种分析物的回收率在74.5%至96.5%之间，变异系数低于16.7%。因此，这种免疫测定法可作为一种快速简便的方法，用于筛选动物源性食品中五种苯二氮䓬类药物的残留量。
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