一种通用型半抗原免疫层析试纸的制备及在双咪苯脲上的应用  

Preparation of Universal Hapten Immunochromatographic Strip and Its Application in Imidocarb

作　　者：邢云瑞 孙亚宁[1] 胡骁飞[1] 范璐[1] 陈琳琳 张改平[1] XING Yunrui;SUN Yaning;HU Xiaofei;FAN Lu;CHEN Linlin;ZHANG Gaiping(Institute for Animal Health(Key Laboratory of Animal Immunology),Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;College of Food Science and Technology,Henan Agricultural University,Zhengzhou 450002,China)

机构地区：[1]河南省农业科学院动物疫病防控研究所(河南省农业科学院动物免疫学重点实验室),河南郑州450002 [2]河南农业大学食品科学技术学院,河南郑州450002

出　　处：《河南农业科学》2024年第9期117-125,共9页Journal of Henan Agricultural Sciences

基　　金：河南省农业科学院自主创新项目(2023ZC086);河南省重点研发与推广专项(科技攻关)(222102110413);河南省农业科学院优秀青年科技基金项目(2022YQ25);国家重点研发计划项目(2018YFC1602903)。

摘　　要：为了提高单克隆抗体筛选效率,降低免疫层析检测方法建立门槛,利用生物素标记人工抗原为探针,以亲和素固定于硝酸纤维素膜上为检测线,建立了通用型半抗原免疫层析试纸,并将其应用于双咪苯脲(IM)单克隆抗体的筛选及残留检测。结果表明,利用碳二亚胺法成功制备了IM人工抗原及生物素探针;通过免疫小鼠及杂交瘤细胞技术制备单克隆抗体;应用试纸筛选,成功获得IM单克隆抗体细胞株1株,其效价为1∶32000,灵敏度为6.25 ng/mL。基于该单抗及通用型半抗原免疫层析试纸建立了IM免疫层析检测试纸,检测牛奶中IM的灵敏度为7 ng/mL。该试纸及其探针制备方法可以通用于半抗原抗体筛选,为抗体筛选提供了一种更高效的方法;无需免疫层析试纸研发平台即可建立半抗原免疫层析检测方法,为免疫层析试纸的研发提供了新的技术手段。In order to improve the selection efficiency of monoclonal antibodies and lower the preparation threshold of immunochromatographic strip,a universal hapten immunochromatographic strip was established by using biotin⁃labeled artificial antigen as a probe and avidin in a test line,and applied for selecting monoclonal antibodies against imidocarb(IM)and detecting IM residues.The results showed that IM artificial antigen and biotin probe were successfully prepared by 1⁃(3⁃dimethylaminopropyl)⁃3⁃ethylcarbodiimide(EDC)method.Subsequently,monoclonal antibodies were prepared by immunizing mice and hybridoma cells technology,and the universal immunochromatographic strip was established.Using the strip,a monoclonal antibody cell line against IM with a titer of 1∶32000 was successfully selected,and its sensitivity was up to 6.25 ng/mL.Based on the excellent monoclonal antibody and the universal strip,an immunochromatographic test strip for the detection of IM was established,and its sensitivity was 7 ng/mL in the milk.The immunochromatographic strip and its probe preparation method can be used in general hapten antibody screening,and a hapten immunochromatographic strip test can be established without professional research and development platform,which provides a more efficient method for antibody screening and a new technology for the development of immunochromatographic strip.

关 键 词：半抗原 通用型免疫层析试纸 双咪苯脲 抗体筛选 残留检测 

分 类 号：R446.6[医药卫生—诊断学]