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本试验制备了用于AFB_1检测的SPR芯片,建立了检测饲料中黄曲霉毒素SPR检测方法,用于玉米样品中AFB_1的快速定量检测。将偶联抗原AFB_1-OVA通过卵清蛋白的氨基酸结合到羧基修饰的IB-CM SPR芯片上,进行表面活化。根据抗原包被的结合区间优化和确定芯片检测条件,确定使用乙酸盐缓冲液、最合适的包被浓度、单抗最适浓度,建立了直接竞争SPR检测法。AFB_1线性区间在0.78-2ppb之间,检测限为0.78ppb。通过实际检测玉米数据分析,SPR法符合正态分布,但与HPLC法检测存在一定的差异。可能与SPR检测方法有关,也可能与样本的不均一性有关。样本中AFB_1浓度越低,表现出的检测差异越大。但使用这种全新的SPR法检测AFB_1,无需标记、实时的检测使得免疫分析更加简单和快速,分析过程可以完全自动化,能避免和减少引入误差。
In this study, an SPR chip for AFB_1 detection was prepared and a method for determination of aflatoxin SPR in feedstuffs was established for the rapid quantitative detection of AFB_1 in maize samples. The conjugated antigen AFB_1-OVA was surface-activated by binding the amino acids of ovalbumin to the carboxy-modified IB-CM SPR chip. Optimize and determine the chip detection conditions based on the antigen-coated binding interval, and establish the direct competition SPR detection method using acetate buffer, the most suitable coating concentration and the optimal concentration of the monoclonal antibody. The linear range of AFB_1 was between 0.78-2ppb and the limit of detection was 0.78ppb. Through the actual analysis of maize data, the SPR method is in accordance with the normal distribution, but there is a certain difference with the HPLC method. May be related to SPR detection methods, may also be related to the sample heterogeneity. The lower the AFB_1 concentration in the sample, the greater the difference in detection. But AFB_1 is detected using this new SPR method, eliminating the need for labeling. Real-time detection makes immunoanalysis simpler and faster, and the analysis process is fully automated, avoiding and reducing the introduction of errors.