作　　者： ; ; ; ; ; ; ; ;

机构地区： 华南农业大学食品学院广东省高等学校食品质量安全重点实验室

出　　处： 《分析化学》 2012年第5期745-751,共7页

摘　　要： 本研究针对呋喃唑酮代谢物(AOZ),设计合成了系列半抗原,进一步通过偶联牛血清白蛋白(BSA)免疫Balb/c小鼠、细胞融合、筛选和亚克隆等过程成功获得了源于新颖半抗原H3的具有高亲和力(亲和力常数6.68×1010 L/mol)和高特异性(与其它功能类似物交叉反应小于0.1%)抗AOZ单克隆抗体。同时,基于设计合成的系列同/异源半抗原/包被抗原,考察了不同结构包被原对ELISA方法灵敏度的影响。另外,采用最佳的特征结构异源包被原H5-OVA,建立了以对硝基苯甲醛(p-NP)为衍生剂的AOZ间接竞争ELISA(icELISA)和直接竞争ELISA(dcELISA)检测方法。结果表明:icELISA模式的AOZ检测IC50为0.503μg/L,定量检测线性范围(IC20～IC80)为0.06～14.0μg/L,检出限(IC10)达0.017μg/L;dcELISA模式的AOZ检测IC50为1.19μg/L,定量检测线性范围为0.14～23.6μg/L,检出限为0.056μg/L。两种方法对AOZ的检测灵敏度和定量线性范围均达到相关检测限量要求,可满足不同需求的实际样品检测。 Several new haptens for furazolidone metabolites 3-amino-2-oxazolidnone（AOZ） were synthesized and coupled to bovine serum albumin（BSA） as immunogen and used to immunize Balb/c mice.A cell strain secreting anti-AOZ monoclonal antibody from hapten H3 was obtained by cell fusion and screening.The titer of antibod was 1∶2000000,the subtype was IgG1 and the affinity constant was equal to 6.68×1010 L/mol.The monoclonal antibody obtained was highly specific to AOZ.Furthermore,based on investigating the effect of homology and heterologous coating antigens with different structures on the ELISA sensitivity,indirect competitive ELISA（icELISA） and direct competitive ELISA（dcELISA） test methods could be established with H5-OVA as the best heterologous coating antigen.The IC50,limit of detection and linear range for AOZ were 1.189 μg/L,0.056 μg/L and 0.14-23.6 μg/L for icELISA and 0.503 μg/L,0.017 μg/L and 0.06-14.0 μg/L for dcELISA,respectively.Both formats of ELISA can achieve related requirement of maximum residue limits（MRLs） for AOZ and provide a potential and alternative rapid immunoassay method to detect AOZ in food.

关 键 词： 呋喃唑酮 单克隆抗体 酶联免疫法 异源包被

领　　域： [农业科学] [农业科学] [农业科学]