赭曲霉毒素A完全抗原的制备及鉴定

Preparation and Identification of Complete Antigen for Ochratoxin A

赭曲霉毒素A(OTA)是一种对人和动物具有广泛毒性的霉菌毒素,为建立OTA在食品和饲料中的快速检测方法,采用活性酯法(NHS)将OTA与牛血清白蛋白(BSA)、鸡卵清白蛋白(OVA)偶联,分别制备了免疫原OTA-BSA和包被原OTA-OVA。紫外扫描和SDS-PAGE鉴定表明,OTA与BSA和OTA与OVA偶联成功,偶联物OTA-BSA和OTA-OVA的偶联比分别为7∶1和4.5∶1。用免疫原OTABSA免疫Balb/c小鼠,制备多克隆抗体,用OTA-OVA包被ELISA板进行检测,小鼠血清中抗OTA的抗体效价最高可达1∶51 200。表明制备的免疫原OTA-BSA具有良好的免疫原性,为OTA单克隆抗体的制备奠定了基础。

Ochratoxin A（OTA）is a mycotoxin which has a wide range of toxicity to human and animals.In order to establish a rapid detection method of OTA in food and cereals,OTA was coupled with BSA and OVA using NHS method to prepare the immunogen OTA-BSA and coating antigen OTA-OVA.UV scanning spectrum and SDS-PAGE methods showed that BSA and OVA were successfully coupled with OTA.The coupling ratio of them were 7∶1and 4.5∶1,respectively.The polyclonal antibody was prepared by immunizing Balb/c mice with OTA-BSA,and the titer was up to 1∶51 200 by ELISA after coating ELISA plate with OTA-OVA.The result showed that the prepared immunogen OTA-BSA had a satisfied immunogenicity and provided a basis for the preparation of OTA monoclonal antibody.