呋喃西林代谢物多克隆抗体制备及酶联免疫吸附分析方法

为了建立检测呋喃西林代谢物(SEM)的酶联免疫吸附分析方法(ELISA),将所设计合成的系列半抗原与牛血清白蛋白(BSA)偶联制备免疫原并免疫新西兰大白兔,筛选获得源于新颖半抗原H3的具有高亲和力、高特异性抗SEM多克隆抗体。同时,基于设计合成的系列同/异源包被抗原,考察了不同结构包被原对ELISA灵敏度的影响,发现H4-OVA作为异源包被原建立SEM的ELISA检测方法可获得最佳的检测效果,结果显示:ELISA方法的半抑制浓度(IC50)为12.37ng/mL;定量检测线性范围(IC20～IC80)为0.439～110.78ng/mL;检测限(IC10)达0.07ng/mL,达到了国内外相关检测限量要求,可应用于实际食品样品检测。

Production and identification of polyclonal antibody detect nitrofurazone metabolite and development of enzyme-linked immunosorbent assay

In order that a method of Enzyme-linked immunosorbent assay(ELISA) method was established to detect nitrofurazone metabolite(SEM),series haptens were synthesized and coupled to bovine serum albumin(BSA) as immunogen which was used to immunize New Zealand white rabbit.An anti-SEM PcAb original from new hapten-H3 was obtained and was highly specific to SEM.Effect of ELISA method sensitivity on the different structure coating antigen(homology/heterologous) was inspected.ELISA detection method against SEM could achieve the best result on account of heterologous feature structure H4-OVA.Results showed:half inhibiting concentration(IC50) was 12.37ng/mL,the quantitative detection linear range(IC20~IC80) was 0.439~110.78ng/mL,and the limit of detection limit(IC10) was 0.07ng/mL for ELISA method,which made it met requirement of relevant detection limit at home and abroad and could be applied in the actual demand of food samples detection.