邻苯二甲酸二环己酯人工抗原的制备研究

目的合成邻苯二甲酸二环己酯半抗原衍生物和合格的人工全抗原,探讨其制备方法。方法通过保留环己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环己酯半抗原衍生物4-氨基邻苯二甲酸二环己酯,产物经通过1HNMR、IR和UV确证了结构。半抗原衍生物再通过重氮化反应与BSA偶联。结果得到的半抗原衍生物4-硝基邻苯二甲酸二环己酯的2个紫外吸收峰分别为:λ1=214nm,λ2=256nm;4-氨基邻苯二甲酸二环己酯的2个紫外吸收峰分别为:λ1=226nm,λ2=288nm。人工全抗原4-氨基邻苯二甲酸二环己酯-BSA,经荧光光谱(λex=307nm,λem=468nm)、体系颜色变化和免疫实验确证偶联成功。抗原结合比为19∶1。人工全抗原通过免疫动物得到了效价高、特异性好、纯度也较高的抗体。结论通过保留环己基结构和在芳环上引入氨基的设计思路可以合成邻苯二甲酸二环己酯的人工全抗原。

Study of dicyclohexyl phthalate on preparation and characterization of artificial antigen

OBJECTIVE: To study the preparation and characterization of hapten and artificial antigen of dicyclohexyl phthalate. METHODS: A dicyclohexyl phthalate hapten (4-amino dicyclohexyl phthalate) was synthesized by introducing amino as a substituent on the aromatic ring and retaining the ester group, and characterized by 1HNMR, IR and UV. The hapten was conjugated to BSA via amino diazotization linkage. RESULTS: Lambda1 = 214nm, lambda2 = 256nm for the UV of dicyclohexyl 4-nitrophthalate and lambda1 = 226nm, lambda2 = 288nm for the UV of dicyclohexyl 4-aminophthalate. Artificial antigen was prepared and tested by fluorescence, and lambda(ex) = 307nm, lambda(em) = 468nm, and the approximate molar ratio of dicyclohexyl 4-aminophthalate to BSA was 19. The product was used as an immunogen, demonstrating that it is suitable for polyclonal antibody production. CONCLUSION: It is a good method for preparation of artificial antigen of dicyclohexyl phthalate by introducing amino as a substituent on the aromatic ring and retaining the ester group. It was suggested that could supply excellent immune antigen for further preparation of antibody and immunoassay to dicyclohexyl phthalate.