呋喃妥因代谢物AHD单克隆抗体的制备及鉴定Preparation and identification of monoclonal antibody against 1-amino-hydantion
王伟萍;赵芸;柳爱春;陈飞东;胡叶军;叶聪莹;
摘要(Abstract)：
目的制备呋喃妥因代谢物AHD单克隆抗体,并进行鉴定。方法利用对羧基苯甲醛合成1-氨基乙内酰胺脲(AHD)的衍生物1-氨基乙内酰脲-4-羧苯基肟(4-CPAHD),通过活性脂法将4-CPAHD与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联,获得免疫抗原4-CPAHD-BSA和包被抗原4-CPAHD-OVA,采用紫外分光光度法检测偶联是否成功。将人工合成抗原4-CPAHD-BSA免疫BALB/c小鼠,通过杂交瘤技术将免疫小鼠脾脏细胞与Sp2/0骨髓瘤细胞融合,筛选能稳定产生抗体的细胞株;通过间接ELISA法检测该细胞株分泌的单克隆抗体的效价、邻硝基-PAHD(2-NPAHD)对单克隆抗体的半数抑制浓度(IC50),并分析抗体的特异性。结果偶联后4-CPAHD-BSA的最大吸收峰有较明显的偏移,表明4-CPAHD与BSA偶联成功,平均偶联比为17.4∶1。制备的单克隆抗体的效价为1∶64 000,2-NPAHD对单克隆抗体的IC50为1.45μg/L,单克隆抗体与同类抗生素及其代谢物无交叉反应。结论制备的AHD单克隆抗体各项指标均较好,为呋喃妥因代谢物免疫检测试剂的研发奠定了基础。
关键词(KeyWords)： 呋喃妥因代谢物;单克隆抗体;酶联免疫吸附测定
基金项目(Foundation): 2013年杭州市重大科技创新项目《动物性农产品质量安全快速检测(药残)胶体金试剂盒研发与应用》(20131812A05)
作者(Authors): 王伟萍;赵芸;柳爱春;陈飞东;胡叶军;叶聪莹;
DOI: 10.13200/j.cnki.cjb.001659
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