双抗体夹心酶联免疫法检测不同样品中的蓖麻毒素

Determination of ricin by double antibody sandwich enzyme-linked immunosorbent assay in different samples

目的应用酶联免疫吸附分析方法(ELISA)检测待测样品中的蓖麻毒素。方法用蛋白G亲和层析柱纯化蓖麻毒素单克隆抗体(4C13,3D74,5E4和5H6),以3D74及辣根过氧化物酶(HRP)标记的4C13建立的双抗体夹心ELISA对含有蓖麻毒素的多种样品进行检测。结果抗蓖麻毒素的单克隆抗体经亲和层析纯化后具有较高的蛋白纯度,应用HRP标记的4C13与3D74建立双抗体夹心ELISA,对于溶解于磷酸缓冲液中的蓖麻毒素标准品的检测灵敏度可达2.5μg·L-1;对于土壤、面粉、牛奶、咸菜汁、雪碧、可乐和腐乳汁中的蓖麻毒素样品检测的灵敏度为2.5～5.0μg·L-1;与磷酸缓冲液样品相比较,含有相同浓度蓖麻毒素的小鼠和人血清样品ELISA的阳性结果明显减弱。结论双抗体夹心酶联免疫法能够有效用于含有蓖麻毒素样品的检测分析。

Objective To develop a double antibody sandwich enzyme-linked immunosorbent assay （ELISA） for the determination of riein. Methods Anti-ricin monoelonal antibodies（4C13, 3D74, 5E4 and 5H6）were purified on Protein G Sepharose 4 Fast Flow Column. Antibody 4C13 labeled with horseradish peroxidase （HRP） was used to establish the ELISA with 3D74 for riein detection. Results The antibodies against riein were highly purified. The double antibody sandwich ELISA was a very sensitive method for the detection of ri- ein containing sample. The riein detection limit could be as low as 2. 5μg·L^-1 in PBST and 2. 5-5 μg·L^-1 in the solution of soil, flour, milk, pickle, Sprite, Cola and preserved beaneurd juice. The absorbance of ELISA for the ricin in mouse serum or human serum sample was lower than that in PBST. Conclusion The double antibody sandwich ELISA is a sensitive method for the analysis of ricin containing sample.