多效唑人工抗原的合成与鉴定

Synthesis and Identification of Stable Artificial Paclobutrazol Antigen

为合成新的、有效的多效唑人工抗原,采用琥珀酸酐法对多效唑小分子进行结构修饰,以获得含羧基的多效唑半抗原。将纯化后的半抗原分别与牛血清白蛋白和卵清蛋白经N-羟基琥珀酰亚胺活泼酯法偶联制备免疫原和包被原。采用质谱、红外光谱、核磁共振氢谱对多效唑半抗原的结构进行鉴定;采用紫外光谱及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进行鉴定。结果显示成功合成出多效唑人工抗原,为其抗体的制备和免疫学方法的构建奠定了前期研究基础。

In order to synthesize a novel and effective artificial paclobutrazol antigen, succinic anhydride method was used to modify the structure of paclobutrazol. Paclobutrazol hapten（PAC-HS） with an active carboxyl group was obtained, and conjugated separately with BSA（bovine serum albumin） and OVA（ovalbumin） by carbodiimide method. Immunogen and coating antigen were obtained and designated as PAC-HS-BSA and PAC-HS-OVA, respectively. The hapten was characterized by mass spectrometry, infrared spectroscopy and 1H nuclear magnetic resonance spectroscopy. Ultraviolet spectrophotometry was applied to determine the artificial paclobutrazol antigen. The coupling ratios of the conjugates were determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF/MS）. The results suggested that paclobutrazol antigen was successfully synthesized, which will greatly facilitate antibody production and immunoassay development.