卡巴氧的代谢物喹喔啉-2-羧酸人工抗原的合成及其多克隆抗体的.pdfVIP

60 2008, Vol. 29, No. 05 食品科学 ※基础研究 卡巴氧的代谢物喹喔啉-2-羧酸人工抗原的 合成及其多克隆抗体的制备 肖甄磊，谢忠良，何计国 * (中国农业大学食品科学与营养工程学院，北京 100083) 摘 要：实验采用碳二亚胺法将卡巴氧代谢物喹喔啉-2-羧酸(QCA)与牛血清白蛋白(BSA)偶联，采用用混合酸酐 法将其与卵清白蛋白(OVA)偶联，分别将其作为免疫原 (QCA-BSA)和包被原(QCA-OVA)进行抗血清的制备及ELISA 实验。经过多次动物免疫及EILSA 实验，结果表明，获得了高效价的特异性抗体。 关键词：卡巴氧；喹喔啉-2- 羧酸；人工抗原；多克隆抗体；间接竞争 ELISA Synthesis and Identification of Antigen of QCA and Preparation and Identification of Its Polyclonal Antibody XIAO Zhen-lei，XIE Zhong-liang，HE Ji-guo* (College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China) Abstract ：The QCA is coupled to BSA as an immunogen by the method of EDC, as well, the coating antigen (QCA-OVA) were produced by the method of MA. The immunogen and coating antigen have been identified by the method of ultraviolet full wavelength scanner. Polyclonal antibodies against QCA have been raised in four rabbits and purified by using a saturated ammonium sulfate method. The sensitivity and cross-reactivity of polyclonal antibody have been demonstrated in ELISA test. The results showed that the QCA antiserum have a very high degree of sensitivity and specificity. Key words：carbadox；quinoxaline-2-carboxylic acid；artificial antigen；polyclonal antibody；indirect competitive ELISA 中图分类号：TS201 文献标识码：A 文章编号：1002-6630(2008)05-0060-05 卡巴氧(carbadox，CDX)是喹喔啉-1, 4-二氧化物的 1.1 材料、试剂与仪器 取代衍生物之一，是最早合成的一种抗菌药物。其作 4只2.5～3kg的健康雄性德国大白兔，由中国农业 用机理是抑制细菌脱氧核糖核酸(DNA)的合成，具有广 大学实验动物中心提供。 谱的抗菌作用，常用于预防和治疗猪痢疾和仔猪肠炎， 喹喔啉-2- 羧酸标准品(纯度＞ 97%)、N- 羟基琥珀 也常用作生长促进剂。但由于卡巴氧具有较强的致突变 酰亚胺(NHS)、弗氏完全佐剂、弗氏不完全佐剂、牛