叶酸人工抗原合成及免疫抗血清的鉴定

Preparation of Folic Acid Artificial Antigen and Identification of Antiserum

合成并鉴定叶酸(FA)人工抗原,通过免疫动物生产亲和性高、特异性强的FA多克隆抗血清。通过活化酯法合成叶酸-牛血清白蛋白(FA-BSA)人工免疫原,通过混合酸酐法制备叶酸-卵清蛋白通过动物免疫(OVA)包被原,并采用紫外扫描对人工抗原进行结构鉴定,通过动物免疫对人工抗原进行生物学鉴定,并得到多抗血清,并使用ELISA对血清进行鉴定。结果表明,半抗原FA分别与载体蛋白BSA及OVA偶联成功,偶联率分别为3.84和4.70,免疫的6只小鼠的效价均在1.28×105以上,且4号小鼠的抗血清抗FA的IC50为14.28ng/mL。此实验成功合成了FA人工抗原,并得到了高效价,高灵敏性的FA多抗血清,为FA单克隆抗体制备及快速检测试剂盒的研制奠定基础。

The purpose of this study waw to synthesize and identify folic acid（FA） artificial antigen and obtain its polyclonal antiserum.The Folic acid immunogen（FA-BSA） and coating antigen（FA-OVA） were synthesized by active ester method and mixed anhydride method.The structure of FA-BSA and FA-OVA was identified by ultraviolet scanning and the the biological identification of FA-OVA was done by mice immunization.At the same time,polyclonal antiserum was obtained and identified by ELISA.The results showed that FA was successfully link to carrier.The animal immunization results showed that titers against FA of six BALB/c mice were above 1.28×10 5 and the IC 50 of No.4 mice was 14.28 ng/mL.The high-titer mice polyclonal antiserum has been developed.FA-BSA and FA-OVA were synthesized and sensitivity FA polyclonal antiserum has been produced in this study.The immunogen of FA and its mice polyclonal antiserum laid a solid foundation for product the monoclonal antibodies and rapid test reagent against FA.