Screening and Identification of DNA Aptamers against T-2 Toxin Assisted by Graphene Oxide

被引：88

作者：

Chen, Xiujuan ^{[1
]}

Huang, Yukun ^{[1
]}

Duan, Nuo ^{[1
]}

Wu, Shijia ^{[1
]}

Xia, Yu ^{[1
]}

Ma, Xiaoyuan ^{[1
]}

Zhu, Changqing ^{[2
]}

Jiang, Yuan ^{[2
]}

Wang, Zhouping ^{[1
]}

机构：

[1] Jiangnan Univ, Sch Food Sci & Technol, Synerget Innovat Ctr Food Safety & Nutr, State Key Lab Food Sci & Technol, Wuxi 214122, Jiangsu, Peoples R China

[2] Jiangsu Entry Exit Inspect & Quarantine Bur, Anim Plant & Food Inspect Ctr, Nanjing 210001, Jiangsu, Peoples R China

来源：

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY | 2014年 / 62卷 / 42期

基金：

中国国家自然科学基金;

关键词：

T-2; toxin; ycotoxin; aptamer; SELEX; graphen oxide (GO); IN-VITRO SELECTION; SECONDARY STRUCTURE; LIQUID-CHROMATOGRAPHY; PCR AMPLIFICATION; HT-2; IMMUNOASSAY; OCHRATOXIN; POLYMERASE; APTASENSOR; STABILITY;

D O I：

10.1021/jf5032058

中图分类号：

S [农业科学];

学科分类号：

09 ;

摘要：

A high-affinity ssDNA aptamer that specifically binds to T-2 toxin was generated by the systemic evolution of ligands by exponential enrichment (SELEX) procedure assisted by graphene oxide (GO). After 10 rounds of selection against T-2 toxin, a highly enriched ssDNA pool was sequenced and the representative aptamers were subjected to binding assays to evaluate their affinity and specificity. Circular dichroism spectroscopy was also used to study the inherent interaction of T-2 toxin and the preferred aptamer Seq.16, which demonstrated a low dissociation constant (K-d) of 20.8 +/- 3.1 nM and excellent selectivity for T-2 toxin. Using the selected aptamer Seq.16 as the recognition element, an aptamer-based fluorescent bioassay was developed for the measurement of T-2 in beer samples with a linear range from 0.5 to 37.5 mu M (R-2 = 0.988) and a limit of detection (LOD) of 0.4 mu M. The results indicate that GOSELEX technology is appropriate for the screening of aptamers against small-molecule toxins, offering a promising application for aptamer-based biosensors.

引用

页码：10368 / 10374

页数：7

共 38 条

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