有机磷农药通用抗原的合成及鉴定

Synthesis and Identification of Generic Antigen for Organophosphorus Pesticides

以二乙基膦酸乙酸作为通用结构半抗原,经活化酯法(NHS-DCC)、碳化二亚胺法(EDC)和混合酸酐法(MAH)合成了3种免疫原,用MAH法合成了1种包被原。人工抗原用紫外分光光度法初步鉴定偶联结果并计算克分子浓度分别为2.846×10-3、2.133×10-3、3.574×10-3、2.584×10-3。后用核磁共振P谱进一步确定偶联情况。偶联成功后免疫Balb/c小鼠制备多克隆抗体,动物实验检测人工抗原的免疫原性,结果表明:NHS-DCC、EDC和MAH法所制得抗血清的最高滴度分别为24000、10000和32000,均获得免疫应答,以半抗原为研究对象建立的间接竞争ELISA法对半抗原的最低检测质量浓度IC10值为1.115×10-5g/L,抑制中浓度IC50值为7.176×10-4g/L。

Diethylphosphono acetic acid was used as a generic hapten, and conjugated to carrier protein by the active ester method, carbodiimide method and mixed-anhydride method. Generic antigen were identified by UV and NMR31P and shown that the molality of 3 immunogen and 1 primordial covering was 2.846×10-3, 2.133×10-3, 3.574×10-3, 2.584×10-3 respectively. BALB/C rabbites were then immunized with three immunogen using different methods for the preparation of antiserum. The titers of antiserum by those three methods had reached 24000, 10000 and 32000 respectively. The CI-ELISA was conducted with the antiserum, the detection limit for thehapten was 1.115× 10-5 g/L and the IC50 was 7.176× 10-4 g/L.