模拟电影
免疫分析
化学
黄曲霉毒素
单克隆抗体
真菌毒素
噬菌体展示
色谱法
抗体
多克隆抗体
检出限
食品科学
生物化学
生物
免疫学
肽
作者
Fengchun Zhao,Yuan Tian,Qiang Shen,Ruxia Liu,Ruirui Shi,Huimin Wang,Zhengyou Yang
出处
期刊:Talanta
[Elsevier]
日期:2018-11-06
卷期号:195: 55-61
被引量:46
标识
DOI:10.1016/j.talanta.2018.11.013
摘要
Mimotopes could replace mycotoxins and their conjugates to develop immunoassay methods. The mimotopes obtained by phage display technology were mainly using monoclonal antibodies or polyclonal antibodies as targets. However, the mimotope of recombinant antibody has not been selected and applied to immunoassay for mycotoxin. The purpose of this study was to prove that an immunoassay for mycotoxin could be developed based on both recombinant antibody and its mimotope. Using aflatoxin B1 (AFB1) as a model system, mimotopes of an aflatoxin nanobody Nb28 were screened by phage display. A rapid magnetic beads-based directed competitive ELISA (MB-dcELISA) was developed utilizing Nb28 and its mimotope ME17. The 50% inhibitory concentration and the detection limit of the MB-dcELISA were 0.75 and 0.13 ng/mL, respectively, with a linear range of 0.24-2.21 ng/mL. Further validation study indicated good recovery (84.2-116.2%) with low coefficient of variable (2.2%-15.9%) in spiked corn, rice, peanut, feedstuff, corn germ oil and peanut oil samples. The developed immunoassay based on nanobody and mimotope provides a new strategy for the monitoring of AFB1 and other toxic small molecular weight compounds.
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