TNT荧光偏振免疫检测方法研究

摘要

One fluorescence polarization immunoassay (FPIA) was developed for detecting 2,4,6-Trinitrotoluene (TNT) based on its polyclonal antibody. Animals were immunized by the synthesized artificial antigen and TNT specific antibody can be obtained. The fluorescence - labeled TNT was synthesized with 1-PyrenyldiazoMethane (PDAM) and purified by solid phase extraction (SPE). The TNT FPIA had a dynamic range from 13. 8 to 129. 7 μg/L with IC50 value of 39. 3 μg/L and the detection limit was 7. 5 μg/L. The FPIA method is of higher sensiticity, simplicity and stability, so it is a potential way to determine TNT.%研究旨在利用新制备的多克隆抗体首次建立2,4,6-三硝基甲苯(TNT)的荧光偏振免疫分析方法.通过设计合成新的TNT半抗原TNT-BSA,偶联载体蛋白后免疫新西兰大白兔,制备亲和力高、特异性强的多克隆抗体.结合新设计合成的荧光示踪物TNT-PDAM建立了TNT的荧光偏振免疫分析方法.试验结果表明,TNT荧光偏振免疫分析方法检测限为7.5μg/L,检测范围为13.8～129.7 μg/L,IC50为39.3μg/L.