Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences , Wuhan 430062, P. R. China.
J Agric Food Chem. 2013 Mar 13;61(10):2426-33. doi: 10.1021/jf4004048. Epub 2013 Feb 27.
To search for an alternative to using protein conjugated aflatoxin as a coating antigen in aflatoxin detection by an ELISA method, a random-8-peptide library was constructed and used as a source of peptides that mimic aflatoxins (termed as mimotopes). Five mimotope peptides were obtained by panning-elution from the library and were successfully used in an indirect competitive ELISA for analyzing total aflatoxin concentration. The assay exhibited an IC50 value of 14 μg/kg in samples (with 1 in 7 dilution of sample extract) for aflatoxins. The linear range is 4-24 μg/kg. Further validation indicated relatively good recovery (60-120%) in peanut, rice and corn. Natural contaminated samples (peanut and feedstuff) were analyzed for aflatoxin concentration by both conventional ELISA and phage ELISA. The results showed good correlation. It can be concluded that the mimotope preparation is an effective substitute for the aflatoxin based coating antigen in ELISA and can be used in real sample analysis.
为了寻找替代物来替代 ELISA 方法中使用的蛋白结合黄曲霉毒素作为包被抗原，构建了一个随机 8 肽库，并将其用作模拟黄曲霉毒素的肽的来源（称为模拟表位）。通过从文库中进行淘选-洗脱，获得了 5 个模拟表位肽，并成功地用于间接竞争 ELISA 分析总黄曲霉毒素浓度。该测定法在样品中的 IC50 值为 14 μg/kg（样品提取物稀释 1/7）。线性范围为 4-24 μg/kg。进一步的验证表明，在花生、大米和玉米中的回收率相对较好（60-120%）。通过常规 ELISA 和噬菌体 ELISA 对天然污染样品（花生和饲料）进行了黄曲霉毒素浓度分析。结果表明相关性良好。可以得出结论，模拟表位制备是 ELISA 中基于黄曲霉毒素的包被抗原的有效替代品，可用于实际样品分析。
