Synthesis and analysis of complete antigen of diethylstilbestrol

WANG Bei; PENG Chifang; XU Chuanlai

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Synthesis and analysis of complete antigen of diethylstilbestrol

更新时间：2025-12-10

- Synthesis and analysis of complete antigen of diethylstilbestrol
- CIESC Journal Issue 6, Pages: 1523-1528(2007)
- 作者机构：
  
  江南大学食品学院
- 作者简介：
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- Online First：05 June 2007，
  
  Published：05 June 2007
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WANG Bei, PENG Chifang, XU Chuanlai. Synthesis and analysis of complete antigen of diethylstilbestrol[J]. CIESC Journal, 2007, (6): 1523-1528.
DOI：

WANG Bei, PENG Chifang, XU Chuanlai. Synthesis and analysis of complete antigen of diethylstilbestrol[J]. CIESC Journal, 2007, (6): 1523-1528. DOI：

摘要

以己烯雌酚（DES）﹑氯乙酸钠(CH2ClCOONa)和牛血清蛋白（BSA）等为主要原料;首先用氯乙酸钠修饰DES分子中的一个酚羟基从而合成单羧甲醚己烯雌酚(DES-MCME);然后以混合酸酐法将DES-MCME与载体蛋白（BSA﹑OVA）偶联形成完全抗原;并以紫外扫描分析完全抗原的偶联比率;再将得到的免疫原DES-BSA按免疫程序免疫新西兰白兔。结果表明己烯雌酚及其衍生物存在着顺反异构转化现象;极性有机溶剂的溶剂化作用促使顺反异构体的转化。高分辨质谱测得合成的DES-MCME相对分子质量为325。人工免疫原及包被原的最适结合比为13和10。采用间接酶联免疫测定;产生的抗血清的稀释度可以达到102400。从而建立了DES-MCME的制备方法;并通过高分辨质谱和ELISA检测方法验证了DES人工抗原的有效性;为研制DES残留免疫检测试剂盒奠定了基础。

Abstract

Diethylstilbestrol monocarboxymethoxyflurane (DES-MCME) was synthesized by first retouching a phenolic hydroxyl group of DES molecule and using diethylstilbestrol;sodium monochloroacetate and bovine serum albumin as key raw materials.The conjugations of DES-MCME with proteins as a complete antigen were prepared by the mixed acid anhydride method and the conjugations were analyzed by ultraviolet scanning.According to the immune protocol;New Zealand white rabbits were immunized.The results of experimentation showed that isomerism was present in cis-and trans-forms of DES and its derivates;which was effected obviously by polar solvents.High resolution mass spectroscopy revealed that the molecular mass of DES-MCME was 325.The optimal conjugation ratios of immunogen and coating antigen were 13 and 10.The dilution rate of antiserum could reach 102400;through an indirect enzyme-linked immunosorbent assay (ELISA).The preparation method of DES-MCME was developed.The effectiveness of the artificial antigen of DES was established by high resolution mass spectroscopy and ELISA methods.This could provide the basis for the DES ELISA kit.

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