沙拉沙星完全抗原的合成及多克隆抗血清制备  

Synthesis of Complete Antigen and Preparation of Polyclonal Antiserum against Sarafloxacin

作　　者：刘儒彪[1,2] 胡骁飞[2] 孙亚宁[2] 王方雨[2] 蔡齐超 邓瑞广[2] 张改平[2,3] 刘玺[1] 

机构地区：[1]河南科技学院食品学院,河南新乡453003 [2]河南省农业科学院河南省动物免疫学重点实验室,河南郑州450002 [3]河南农业大学牧医工程学院,河南郑州450002

出　　处：《河南农业科学》2014年第5期167-171,共5页Journal of Henan Agricultural Sciences

基　　金：国家科技支撑计划项目(2011BAK10B01);河南省重大科技专项项目(121100110800);河南省高校科技创新团队支持计划项目(13IRTSTHN006);公益行业(农业)科研专项(201203040)

摘　　要：为了制备沙拉沙星(SARA)多克隆抗血清,采用碳二亚胺(EDC)法将SARA分别与牛血清白蛋白(BSA)、卵清白蛋白(OVA)偶联,制备了免疫原SARA-BSA、包被原SARA-OVA。经SDS-PAGE电泳与紫外扫描初步判定偶联成功,然后用免疫原SARA-BSA分别按20、50μg/只的剂量免疫BALB/c小鼠,共免疫3次,每次间隔3周,于最后1次免疫10d后断尾采血,制备多克隆抗血清。采用间接ELISA方法测定多克隆抗血清效价,间接竞争ELISA方法测定其敏感性和特异性。结果显示,制备多克隆抗血清效价均达1∶10 000以上,IC50为623.5ng/mL,且与其他药物交叉反应率低,具有较好的特异性。To prepare high sensibility and specificity polyclonal antiserum against Sarafloxacin （SARA）,SARA was linked to the carrier proteins BSA and OVA with EDC method to prepare the immunogen SARA-BSA and the coating antigen SARA-OVA.UV scanning spectrum and SDS-PAGE preliminarily proved that SARA was coupled successfully to the carrier protein.The specific polyclonal antiserums with higher titer were obtained by immunizing BALB/c mice with SARA-BSA of 20 μg,50 μg each mouse respectively,with the titer of 1 ∶ 10 000 and the inhibition titer of 623.5 ng/mL.These results indicated that SARA was successfully linked to the carrier proteins.This study laid the foundation for further gain monoclonal anti-SARA and its immunological assay.

关 键 词：沙拉沙星 人工抗原 ELISA 多抗血清 

分 类 号：S859.84[农业科学—临床兽医学]