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喹喔啉-2-羧酸人工抗原合成与抗体制备
引用本文:张泽英,袁宗辉.喹喔啉-2-羧酸人工抗原合成与抗体制备[J].中国畜牧兽医,2013,40(10):152-155.
作者姓名:张泽英  袁宗辉
作者单位:1. 武汉生物工程学院食品工程系, 湖北武汉 430015;2. 农业部食品安全评价重点开放实验室, 湖北武汉 430070
基金项目:上海市科技兴农计划(沪农科攻字2003B99)。
摘    要:试验以喹喔啉-2-羧酸(quinoxaline-2-carboxylic acid,QCA)及其与γ-氨基丁酸(ABA)的衍生物(QCA-ABA)为半抗原,将其与牛血清白蛋白(BSA)和鸡卵清蛋白(OVA)偶联,合成人工抗原。以QCA-BSA和QCA-ABA-BSA为免疫原,免疫大白兔制备多克隆抗体,利用紫外分光光度计分析结合物的结合情况,间接ELISA和间接竞争ELISA分析抗体特性。结果表明,以QCA-BSA、QCA-ABA-BSA为免疫原制备的抗体平均效价分别为12.8×104、6.4×104。2种抗体对QCA灵敏度低,IC50均大于10000 ng/mL,而对QCA-ABA的灵敏度高,IC50分别为14.2和7.75 ng/mL,特异性好,仅与3-甲基-喹喔啉-2-羧酸(12%)、卡巴氧(0.038%)和喹赛多(0.028%)有较低程度的交叉反应,与衍生试剂和结构类似物均无交叉反应(<0.01%)。该抗体为建立动物可食性组织中QCA残留物的快速检测方法奠定了基础。

关 键 词:喹喔啉-2-羧酸  卡巴氧  半抗原  人工抗原  抗体  
收稿时间:2012-09-11

Antigens Synthesis and Antibodies Preparation of Quinoxaline-2-Carboxylic Acid
ZHANG Ze-ying,YUAN Zong-hui.Antigens Synthesis and Antibodies Preparation of Quinoxaline-2-Carboxylic Acid[J].China Animal Husbandry & Veterinary Medicine,2013,40(10):152-155.
Authors:ZHANG Ze-ying  YUAN Zong-hui
Institution:1. Department of Food Engineering, Wuhan Bioengineering Institute, Wuhan 430015, China;2. MOA Key Laboratory of Food Safety Evaluation, Wuhan 430070, China
Abstract:Quinoxaline-2-carboxylic acid (QCA) and γ-amino butyric acid derivative of QCA (QCA-ABA) were conjugated to bovine serum albumin (BSA). The conjugates QCA-BSA, QCA-ABA-BSA used to immunize New Zealand White rabbits were identified with UV scanning spectrum and the specificity of antibodies were detected by indirect ELISA and indirect compete ELISA. The antibodies produced with QCA-BSA, QCA-ABA-BSA conjugates showed the titers of 12.8×104, 6.4×104, respectively. These antibodies were not specific for the QCA moiety, IC50 values of >10000 ng/mL, but specific for the QCA derivatization (QCA-ABA), IC50 values of 14.2 and 7.75 ng/mL, respectively, and cross-reactivity of 12% with 3-methyquinoxaline-2-carboxylic acid, 0.038% with carbadox and 0.028% with cyadox, but did not cross-react with other veterinary drugs. The study laid a solid foundation for rapid detection of QCA in edible animal tissues.
Keywords:quinoxaline-2-carboxylic acid  carbadox  hapten  artificial antigen  antibody  
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