本研究为了克服现有SEM半抗原结构不含硝基端基团的不足，将SEM与4-（3-醛基-4-硝基-苯氧基）-丁酸衍生制备半抗原，保留了硝基端基团。然后，将制备的半抗原分别与BSA和OVA偶联制备SEM-BSA免疫原和SEM-OVA包被原。免疫原免疫BALB/C小鼠后，结合细胞融合技术制备单克隆抗体，并利用间接竞争酶联免疫分析法（ciELISA）评价单克隆抗体的效价及特异性。结果显示，单克隆抗体可特异性识别SEM的2-硝基苯甲醛衍生物（2-NPSEM），效价达1:2560000。应用ciELISA检测法，在0.05~4.05 μg/L呈线性关系，检测限高达0.04 μg/L，IC50值为0.23 μg/L，与结构类似物NPAMOZ、NPAOZ和NPAHD的交叉反应率分别为0.03%、0.02%和0.26%。本论文提供的SEM半抗原的设计和改造方法，为呋喃西林的代谢物SEM的高效检测提供了新思路和方法。

In the present work,a novel method for preparation of nitro-terminal group-containing SEM hapten was established by a reaction of SEM with 4-(3-aldehyde-4-nitro-phenoxy)-butyric acid s, which introduced a reactive terminal nitro groups into SEM structure. The prepared SEM hapten was then coupled with bovine serum albumin (BSA) and ovalbumin (OVA) respectively to prepare SEM-BSA immunogen and SEM-OVA coating antigen. The BALB/C mice were immunized with immunogen SEM-BSA, and the monoclonal antibodies were prepared by cell cell fusion technology. The titer and specificity of the monoclonal antibodies were evaluated by using indirect competitive enzyme-linked immunoassay (ciELISA). The results showed that the prepared monoclonal antibody could specifically recognize the 2- nitrobenzaldehyde derivates (2-NPSEM) of SEM with a titer of 1:2560000. The established ciELISA method exhibited a good linear relationship in the range of 0.05~4.05 μg/L with a detection limit of 0.04 μg/L and a half maximal inhibitory concentration value (IC50) of 0.23 μg/L, and the cross reaction rates of NPAMOZ, NPAOZ and NPAHD were 0.03%, 0.02% and 0.26%, respectively. This methods for SEM hapten synthesis provided new ideas and ways for the high-efficient detection of nitrofurazone metabolite.

国家自然科学基金资助项目（31271889）