Application of Mimotope Peptides of Fumonisin B1 in Peptide ELISA
Application of Mimotope Peptides of Fumonisin B1 in Peptide ELISA
复制标题
伏马菌素b1模拟表位肽在肽ELISA中的应用。
DOI：
10.1021/jf400056p
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发表时间：
2013-05-22
影响因子：
6.1
通讯作者：
Xiong, Zheng-ping
中科院分区：
文献类型：
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作者：
Liu, Xing;Xu, Yang;Xiong, Zheng-ping
以抗伏马菌素B-1（FB 1）单克隆抗体B 1D 11为靶，从噬菌体随机环约束七肽库中筛选出具有特异性的七肽。在三个淘选循环后，选择具有三个模拟表位肽的七个引物来模拟FB 1与1D 11的结合。经噬菌体ELISA鉴定，选择检测线性范围最好的噬菌体克隆进行进一步研究。合成了一种具有噬菌体的插入肽序列的肽，命名为CT-452。以CT-452-牛血清白蛋白偶联物为包被抗原，建立了检测FB 1的间接竞争ELISA（肽ELISA）。抑制曲线的线性范围为1.77-20.73 ng/mL。半抑制浓度（IC_（50））为6.06 ng/mL，检测限为1.18 ng/mL。将该方法与传统的间接ELISA（商品化ELISA试剂盒）和高效液相色谱（HPLC）进行了比较，结果表明该方法用于谷物样品中FB 1检测的可靠性。评价了肽ELISA中CT-452和FB 1标准品浓度之间的关系。结果表明，合成肽CT-452可以取代FB 1标准品，建立不含FB 1的免疫测定法。
Anti-fumonisin B-1 (FB1) McAb 1D11 was used as the target for biopanning from a phage random loop-constrained heptapeptide library. After three cycles of panning, seven phages with three mimotope peptides were selected to mimic the binding of FB1 to 1D11. After the identification of phage ELISA, the phage clone that showed the best linear range of detection was chosen for further research. One peptide with the inserted peptide sequence of the phage was synthetized, named CT-452. An indirect competitive ELISA (peptide ELISA) for detecting FB1 was established using the CT-452-bovine serum albumin conjugate as coating antigen. The linear range of the inhibition curve was 1.77-20.73 ng/mL. The half inhibitory concentration (IC50) was 6.06 ng/mL, and the limit of detection was 1.18 ng/mL. This method was compared with conventional indirect ELISA (commercial ELISA kit) and high-performance liquid chromatography (HPLC), and the results showed the reliability of the peptide ELISA for the determination of FB1 in cereal samples. The relationship between the CT-452 and FB1 standard concentrations in peptide ELISA was evaluated. The results indicated that synthetic peptide CT-452 can replace the FB1 standard to establish an immunoassay free of FB1.