食品工业科技2013，Vol.34Issue(13)：161-165,5.
呋喃它酮代谢物5-甲基吗啉-3-氨基-2-恶唑烷酮单克隆抗体的制备、鉴定与胶体金免疫层析试纸条的研制
Production and characterization of monoclonal antibody against the metabolite of furaltadone,5-morpholino-3-amino-2-oxazolidone (AMOZ), and establishment of colloidal gold immunochromatography for detecting
摘要
Abstract
Object:The monoclonal antibody against 5-morpholino-3-amino-2-oxazolidone(AMOZ)was prepared,and the corresponding colloidal gold immunochromatography method was established.Method:The hapten CP-AMOZ was synthesized by the reaction between AMOZ and 4-CBA and conjugated with BSA and OVA using activated ester method.Ascitic type antibody was prepared by hybridom cell lines which secret monoclonal antibody against CP-AMOZ by mean of immunizing BALB/c mice,cell fusion,ELISA screening and subcloning.Data including the titer of antibody,its IC50 against CP-AMOZ and LOD were detected by ELISA analysis and determine its specificity.Competitive colloidal gold immunochromatographic dipstick was produced on this base,gold nanoparticles were coupled with monoclonal antibodies,CP-AMOZ-BSA and rabbit-anti-mouse polyclonal antibody were separately coated on nitrocellulose membrane as test line and control line to detect the limit of detection and specificity.Result:The titer of the mAbs was 1∶1.6 x 105,the IC50 value and LOD to CP-AMOZ were 0.86μg/L and 0.07μg/L respectively.The mAbs had little cross reactivity with other nitrofurans metabolites and derivates,showing high specificity to CP-AMOZ,and LOD of colloidal gold immunochromatographic dipstick was 5μg/LConclusion:The antigen of CP-AMOZ was successfully synthetized and the monoclonal antibody specific to CP-AMOZ was prepared,which help prepare the colloidal gold rapid detection dipstick of high sensitivity and specificity.关键词
CP-AMOZ/单克隆抗体/胶体金/免疫层析Key words
CP-AMOZ/ monoclonal antibody/ colloidal gold/ immunochromatography分类
生物科学引用本文复制引用
张晓丽,颜露,向军俭,邹军辉,唐勇..呋喃它酮代谢物5-甲基吗啉-3-氨基-2-恶唑烷酮单克隆抗体的制备、鉴定与胶体金免疫层析试纸条的研制[J].食品工业科技,2013,34(13):161-165,5.基金项目
广东省教育部"产学研"结合项目(2009B090300452). （2009B090300452）
