呋喃唑酮代谢物AOZ的抗体制备及初步应用  

Preparation and application of monoclonal antibody against furazolidone metabolite residues AOZ

作　　者：李子颖 郑婷 袁志刚 刘一兵 韩世泉[3] 

机构地区：[1]原子高科股份有限公司,北京102413 [2]北京市公安司法医鉴定中心,北京100192 [3]中国原子能科学研究院,北京102413

出　　处：《食品科技》2013年第4期309-312,316,共5页Food Science and Technology

摘　　要：制备呋喃唑酮代谢物AOZ的单克隆抗体并建立AOZ检测的酶联免疫分析方法。用CPAOZ-BSA免疫Balb/c小鼠,取免疫小鼠的脾细胞与Sp2/0融合,经过筛选和克隆化后得到5株可分泌抗AOZ衍生物NPAOZ的单克隆抗体的杂交瘤细胞株。其中CPAOZ45D7对NPAOZ的抑制效果最好。经鉴定,CPAOZ45D7的亲和常数为2.5×109L/mol。用CPAOZ45D7包被酶标板,CPAOZ-HRP作为酶标记物建立了检测AOZ的酶联免疫分析,其曲线范围为1.3～130.0ng/mL,灵敏度为0.2ng/mL,批内变异<15%,批间变异<15%。用该方法测得AOZ在牛奶中的回收率为88%～121%Monoclonal antibody against furazolidone metabolite residues AOZ was preparaed and an direct competitive enzyme-linked immunosobent assay（ELISA） for AOZ was developed.Splenocytes from mice immunized with CPAOZ-BSA were fused with sp2/0 myeloma cells,5 stable monoclonal antibodies against NPAOZ which was the derivative of AOZ were got after selecting and cloning.CPAOZ45D7 has the best inhibitive result against NPAOZ and its affinity constant was 2.5×109 L/mol.An direct competitive ELISA was developed by Coating CPAOZ45D7 and labling CPAOZ with HRP.The standard curve ranged from 1.3 ng/mL to 130.0 ng/mL,the limit of detection for AOZ was 0.2 ng/mL,the intra-and intro-assay CVs were less than 15%,the average recovery of AOZ in milk was 88%～121%.

关 键 词：AOZ 单克隆抗体 酶联免疫分析 

分 类 号：TS207.5[轻工技术与工程—食品科学]