Zhao Qiuling, Zhang Zhen, Xu Li, Xia Ted, Li Nan, Liu Jianli, Fang Xiaohong
Beijing National Laboratory for Molecular Science, Key Laboratory of Molecular Nanostructures and Nanotechnology, Institute of Chemistry, Chinese Academy of Sciences, Beijing, 100190, China.
Anal Bioanal Chem. 2014 May;406(12):2949-55. doi: 10.1007/s00216-014-7705-z. Epub 2014 Mar 6.
A novel enzyme-linked aptamer assay (ELAA) with the aid of Exonuclease I (Exo I) for colorimetric detection of small molecules was developed. The fluorescein isothiocyanate (FITC)-labeled aptamer was integrated into a double-stranded DNA (dsDNA). In the presence of target, the binding of aptamer with target protected the aptamer from Exo I degradation, which resulted in the FITC tag remaining on the aptamer. Then, the anti-FITC-HRP conjugate was used to produce an optically observable signal. By monitoring the color change, we were able to detect two model molecules, ATP and L-argininamide, with high selectivity and high sensitivity even in the serum matrix. It is expected to be a simple and general ELAA method with wide applicability.
开发了一种借助核酸外切酶I（Exo I）用于比色检测小分子的新型酶联适体分析方法（ELAA）。异硫氰酸荧光素（FITC）标记的适体被整合到双链DNA（dsDNA）中。在存在靶标的情况下，适体与靶标的结合保护适体不被Exo I降解，这导致FITC标签保留在适体上。然后，使用抗FITC-HRP偶联物产生可光学观察到的信号。通过监测颜色变化，我们能够在血清基质中高选择性和高灵敏度地检测两种模型分子，ATP和L-精氨酰胺。预计这将是一种具有广泛适用性的简单通用的ELAA方法。
