为合成新的、有效的氯吡脲(forchlorfenuron,CPPU)人工抗原,在无水三氯化铝的催化作用下,采用傅克反应对CPPU小分子进行结构改造。采用碳二亚胺(carbodiimide,EDC)法将半抗原分别与牛血清白蛋白(bovine serum albumin,BSA)和卵清蛋白(ovalbumin,OVA)进行偶联,获得CPPU的免疫抗原(CPPU-BSA)和包被抗原(CPPU-OVA)。采用质谱、元素分析和核磁共振氢谱对CPPU半抗原(CPPU-COOH)的分子结构式进行鉴定;采用紫外光谱和高性能基质辅助激光解吸电离-飞行时间质谱对偶联物的结构和相对分子质量进行鉴定。结果显示成功合成出CPPU人工抗原,为其抗体的制备和免疫学方法的构建提供了参考。
This work aimed to develop an effective novel method to synthesize artificial antigens of forchlorfenuron(CPPU).Towards this goal,Friedel-Crafts reaction was used to modify the structure of forchlorfenuron with the catalysis of anhydrous aluminum chloride and the hapten was coupled with bovine serum albumin(BSA) and ovalbumin(OVA)by carbondiimine(EDC) method,respectively,to obtain forchlorfenuron immunogen and coating antigen.The molecular structure of the forchlorfenuron hapten was identified by elemental analysis,mass spectroscopy(MS) and nuclear magnetic resonance spectroscopy(~1H-NMR).The artificial antigens were identified by UV spectroscopy and high performance matrix assisted laser desorption ionization time of flight mass spectroscopy(MALDI-TOF-MS).The experimental results showed that the artificial antigen of forchlorfenuron has been synthesized successfully and it can lay the foundation for the preparation of antibody and the development of immunoassays.