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当前位置: 首页 > 文章 > 沙拉沙星完全抗原的合成及多克隆抗血清制备 河南农业科学 2014,43 (5) 167-171
Position: Home > Articles > Synthesis of Complete Antigen and Preparation of Polyclonal Antiserum against Sarafloxacin Journal of Henan Agricultural Sciences 2014,43 (5) 167-171

沙拉沙星完全抗原的合成及多克隆抗血清制备

作  者：
刘儒彪;胡骁飞;孙亚宁;王方雨;蔡齐超;邓瑞广;张改平;刘玺
单  位：
河南科技学院食品学院;河南省农业科学院河南省动物免疫学重点实验室
关键词：
沙拉沙星;人工抗原;ELISA;多抗血清
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。
译  名：
Synthesis of Complete Antigen and Preparation of Polyclonal Antiserum against Sarafloxacin
作  者：
LIU Ru-biao;HU Xiao-fei;SUN Ya-ning;WANG Fang-yu;CAI Qi-chao;DENG Rui-guang;ZHANG Gai-ping;LIU Xi;College of Food Science and Technology,Henan Institute of Science and Technology;Henan Provincial Key Laboratory of Animal Immunology,Henan Academy of Agricultural Sciences;Animal Husbandry and Veterinary College,Henan Agricultural University;
关键词：
Sarafloxacin;;artificial antigen;;ELISA;;polyclonal antiserum
摘  要：
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 20μg,50μg each mouse respectively,with the titer of 1∶10 000and the inhibition titer of 623.5ng/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.

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