适体
玉米赤霉烯酮
指数富集配体系统进化
真菌毒素
色谱法
检出限
化学
生物传感器
离解常数
寡核苷酸
计算生物学
核酸
鉴定(生物学)
生物
分子生物学
生物化学
食品科学
核糖核酸
基因
受体
作者
Xiujuan Chen,Yukun Huang,Nuo Duan,Shijia Wu,Xiaoyuan Ma,Yu Xia,Changqing Zhu,Yuan Jiang,Zhouping Wang
标识
DOI:10.1007/s00216-013-7085-9
摘要
Zearalenone (ZEN) is a nonsteroidal estrogenic mycotoxin produced by Fusarium graminearum on maize and barley. Because most current methods of ZEN detection rely on the use of low-stability antibodies or expensive equipment, we sought to develop a rapid, low-cost determination method using aptamers instead of antibodies as the specific recognition ligands. This work describes the isolation and identification of single-stranded DNA (ssDNA) aptamers recognizing ZEN using the modified systematic evolution of ligands by exponential enrichment methodology based on magnetic beads. After 14 rounds of repeated selection, a highly enriched ssDNA library was sequenced and 12 representative sequences were assayed for their affinity and specificity. The best aptamer, 8Z31, with a dissociation constant (K
d) of 41 ± 5 nM, was successfully applied in the specific detection of ZEN in binding buffer and in real samples based on a magnetic separation/preconcentration procedure. This analytical method provided a linear range from 3.14 × 10−9 to 3.14 × 10−5 M for ZEN, and the detection limit was 7.85 × 10−10 M. The selected aptamers are expected to be used in the potential development of affinity columns, biosensors, or other analytical systems for the determination of ZEN in food and agricultural products.
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