为研究建立有机磷农药倍硫磷的免疫分析法，合成了1种倍硫磷半抗原和2种人工抗原。采用改进的方法将三氯硫磷与3-甲基-4-甲硫基苯酚、β-丙氨酸反应制备了倍硫磷半抗原，产物经薄层层析、1HNMR和质谱鉴定为预期产物。分别将半抗原以活泼酯法、混合酸酐法与BSA、OVA偶联制备了人工抗原BZB和BGO，通过紫外光谱鉴定证明偶联成功。用TNBS比色法测定了BZB、BGO与载体蛋白的克分子结合比，分别为29:1和12:1。以BZB为免疫原免疫Balb/c小鼠，以BGO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倍硫磷的特异性抗血清，从而进一步证明抗原合成成功。本方法为研究建立倍硫磷的免疫分析方法奠定了基础。

In order to develop immunoassays to fenthion, an organophosphorus pesticide, one fenthion hapten and two artificial antigens were synthesized before antibody production. The fenthion hapten was produced by combining thiophosphoryl chloride with 3-methyl-4- methylthiophenol and β-alanine. The production was identified through TLC, 1H-NMR and MS. The hapten was conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) by the active ester method and the mixed anhydride method, respectively. The products, named as BZB and BGO, 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 showed that the binding rates of BZB and BGO came to 29:1 and 12:1, respectively. Balb/C mice were immunized with BZB and competitive indirect enzyme-linked immunosorbent assay (ELISA) indicated the existence of specific antibodies in the antisera which further verified the success of synthesis.

国家自然科学基金项目（30901005）；广东省食品安全应急检测技术研究中心项目（2010A032000001-4）；广东省教育部产学研结合项目（2011A090200029、2010A090200084）