甲胺磷衍生物的合成及人工抗原的制备

食品科学 ›› 2006, Vol. 27 ›› Issue (12): 377-380.

甲胺磷衍生物的合成及人工抗原的制备

肖治理，孙远明，张木明，吴青，雷红涛

华南农业大学食品学院广东省教育厅食品质量安全重点实验室；

出版日期:2006-12-15 发布日期:2011-11-23

Synthesis of Methamidophos Derivative and Preparation of Artificial Antigens

XIAO Zhi-li,SUN Yuan-ming,ZHANG Mu-ming,WU Qing,LEI Hong-tao

College of Food Science, South China Agricultural University, Key Laboratory of Food Quality and Safety of the Education Department of Guangdong Province, Guangzhou 510642, China

Online:2006-12-15 Published:2011-11-23

摘要/Abstract

摘要： O,S-二甲基硫代磷酰氯与β-丙氨酸反应,得到一种甲胺磷的结构衍生物,经薄层层析和1HNMR鉴定,推测预期产物生成。分别采用活泼酯法和混合酸酐法将产物与BSA、OVA偶联制备了人工抗原MZB和MGO,通过紫外光谱鉴定证明偶联成功。用TNBS比色法测定了MZB、MGO与载体蛋白的克分子结合比,分别为16:1和9:1。以MZB为免疫原免疫Balb/c小鼠,以MGO为包被抗原对小鼠血清进行间接竞争ELISA分析表明,小鼠经免疫后产生了针对甲胺磷的特异性抗体,抗血清效价为51:200,从而进一步证明抗原合成成功。本方法简便易行,为研究建立甲胺磷的免疫分析方法奠定了基础。

关键词: 甲胺磷, 衍生, 人工抗原, 抗血清, 免疫分析

Abstract: A methamidophos derivative was synthesized by combining O,S-dimethylthiophosphoryl chloride with β-alanine. The productions were identified through TLC and 1HNMR.The derivative was conjugated to bovine serum albumin and ovalbumin by the active ester method and the mixed anhydride method, respectively. The productions were named MZB and MGO, and were identified through ultraviolet spectrum (UV). A spectrophotometric method using 2,4,6-trinitrobenzene sulfonic acid (TNBS) was performed to estimate hapten to carrier protein ratio. The results came to 16:1 and 9:1 for MZB and MGO respectively. Balb/C mice were immunized with MZB and competitive indirect enzyme-linked immunosorbent assay (ELISA) indicates the existence of specific antibodies in the antisera. The titer of antisera is 51:200. This had further verified the success of synthesis. This study lays the groundwork for further research of immunoassay to methamidophos.

Key words: methamidophos, derivation, artificial antigen, antiserum, immunoassay

肖治理，孙远明，张木明，吴青，雷红涛. 甲胺磷衍生物的合成及人工抗原的制备[J]. 食品科学, 2006, 27(12): 377-380.

XIAO Zhi-li,SUN Yuan-ming,ZHANG Mu-ming,WU Qing,LEI Hong-tao. Synthesis of Methamidophos Derivative and Preparation of Artificial Antigens[J]. FOOD SCIENCE, 2006, 27(12): 377-380.

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