丁香酚半抗原及人工抗原的合成与鉴定

目的 合成丁香酚半抗原及人工抗原并对其结构进行鉴定。方法 采用氯乙酸对丁香酚进行结构改造, 以获得含羧基的丁香酚半抗原。将经重结晶纯化后的半抗原与牛血清白蛋白(bovine serum albumin, BSA)经碳化二亚胺(carbide dimethylamine, EDC)偶联制备人工抗原。采用电喷雾串联三重四极杆质谱法对丁香酚半抗原的结构进行鉴定。采用紫外光谱及基质辅助激光解吸电离-飞行时间质谱(matrix assisted laser desorption ionization-flight time mass spectrometry, MALDI-TOF MS)对偶联物的结构进行鉴定。结果 紫外光谱结果表明, 与载体蛋白相比, 偶联物的特征吸收峰强度有一定的增强; MALDI-TOF MS结果表明, 表明丁香酚半抗原与BSA的偶联比为12.5:1。结论 本研究成功建立了丁香酚半抗原及人工抗原的制备方法, 为丁香酚抗体的制备提供了前期研究基础。

Synthesis and identification of hapten and artificial antigen of eugenol

Objective To synthesize the eugenol haptens and artificial antigens and identify their structures. Methods The structure of eugenol was modified by chloroacetic acid in order to obtain the carboxylated eugenol hapten. Artificial antigen was prepared by combining the recrystallized hapten with bovine serum albumin (BSA) via carbon dimethylamine (EDC). The structure of eugenol hapten was identified by electrospray ionization-triple quadrupole mass spectrometry (ESI-MS/MS). The structure of artificial antigen were identified by ultraviolet (UV) spectrophotometry and matrix assisted laser desorption ionization-flight time mass spectrometry (MALDI-TOF MS). Results The results of UV spectrum indicated that the characteristic absorption peak strength of the conjugate was increased compared with the carrier protein. The results of MALDI-TOF MS revealed that the molar ratio (MR) values of eugenol hapten to BSA was 12.5:1. Conclusion This study successfully establishes the preparation method of eugenol hapten and artificial antigen, which will greatly facilitate antibody production.