邻苯二甲酸二丁酯免疫检测 Ⅰ.人工抗原的制备及表征

Immunological Detection of Dibutyl Phthalate Ⅰ.Preparation and Characterization of Artificial Antigen

通过保留丁酯基结构和在芳环上引入氨基取代基合成了一种邻苯二甲酸二丁酯半抗原衍生物4-氨基邻苯二甲酸二丁酯,通过1H NMR、IR和UV确证了结构,其中4-硝基邻苯二甲酸二丁酯的2个紫外吸收峰分别为:λ1=212 nm,λ2=258 nm;4-氨基邻苯二甲酸二丁酯的2个紫外吸收峰分别为:λ1=226 nm,λ2=288 nm。半抗原衍生物通过重氮化反应与BSA偶联,得到人工抗原,经荧光光谱(λex=307 nm,λem=468 nm)、体系颜色变化和免疫实验确证偶联成功,抗原结合比为13∶1,为进一步的抗体制备和免疫分析提供可靠的免疫原。

Dibutyl phthalate hapten （ dibutyl 4-aminophthalate） was synthesized by introducing amino group as a substituent at the aromatic ring while retaining the ester group. The product was characterized by means of 1^H NMR, IR and UV, with λ1 = 212 nm, λ2 = 258 nm for the UV of dibutyl 4-nitrophthalate and λ1 = 226 nm, λ2 = 288 nm for the UV of dibutyl 4-aminophthalate. The hapten was conjugated to BSA v/a amino diazotization linkage. Thus artificial antigen was prepared and tested by fluorescence spectrum with λ ex = 307 nm, λem =468 nm, and the approximate molar ratio of dibutyl 4-aminophthalate to BSA was 13. The product was used as an immunogen, demonstrating that it is suitable for polyclonal antibody production. The present research has supplied an excellent immune antigen for further preparation and immunoassay of antibody obtained from dibutyl phthalate.