呋喃唑酮人工抗原的制备与鉴定

Synthesis and Characterization of Antigen of Furazolidone

DOI：10.3969/j.issn.1673-1689.2013.11.003

中文关键词: 呋喃唑酮 酶联免疫检测 抗原 抗体

英文关键词: furazolidone ELISA antigen antibody

基金项目:国家“十二五”科技支撑计划项目(2012BAC01B07,2012BAD29B05)

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中文摘要:

为了制备3-氨基-2-恶唑烷酮(AOZ)的完全抗原,作者以乙醇肼和碳酸二甲酯为原料,首先合成了3-氨基-2-恶唑烷酮(AOZ),然后通过对醛基苯甲酸对AOZ进行衍生制得3-(4-羧基苯亚甲基)-氨基-2-恶唑烷酮(CPAOZ),以CPAOZ作为半抗原,经水溶性碳化二亚胺法(EDC),缩合酯化法(DCC)和混合酸酐法(MA)法3种方法分别与载体蛋白偶联,合成人工抗原。对于制备产物通过紫外光谱扫描分析进行了鉴定,3种抗原的偶联比率分别为8、12、17。采用3种抗原做为包被原,建立了酶联免疫(ELISA)抑制曲线。结果表明,对AOZ的半数抑制率(IC50)分别为12.9、7.6、1.1 ng/mL。采用混合酸酐法(MA法)制备的包被抗原与抗体具有较好的匹配性。3-氨基-2-恶唑烷酮(AOZ),是硝基呋喃类抗生素呋喃唑酮在动物体内的稳定代谢物,采用本试验制备的抗原,有望获得特异性更好的抗体。

英文摘要:

In order to prepare complete antigen against 3-amino-2-oxazolidinone(AOZ),which is one of main metabolites of furazolidone in animal,AOZ was synthesized using hydrazine and dimethyl carbonate. Then,3-(4-carboxy benzyl methylene)-amino-2-oxazolidinone(CPAOZ) was prepared as hapten based on AOZ. Three different methods including water-soluble carbodiimide method(EDC),condensation esterification(DCC) and mixed anhydride method(MA) method were used to prepare complete antigens by coupling CPAOZ to ovalbumin(OVA). All the products were characterized with UV spectrum. The coupling ratios of three artificial antigens were respectively 8、12、17. Under the optimized conditions,the prepared antigens were used to establish inhibition curves of AOZ and CPAOZ by enzyme-linked immunoassay(ELISA). Results showed that the lowest half inhibition concentration(IC50) of AOZ was 1.1 ng/mL sourced from coating antigen prepared using mixed anhydride method(MA) method.

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