适体
化学
荧光
共轭体系
配体结合分析
生物物理学
色谱法
生物传感器
阳离子聚合
组合化学
聚合物
生物化学
分子生物学
受体
有机化学
物理
生物
量子力学
作者
Pengbo Zhang,Ke Qin,Anand Lopez,Zhengping Li,Juewen Liu
出处
期刊:Analytical Chemistry
[American Chemical Society]
日期:2022-10-25
卷期号:94 (44): 15456-15463
被引量:17
标识
DOI:10.1021/acs.analchem.2c03564
摘要
With more and more new aptamers being reported, a general, cost-effective yet reliable aptamer binding assay is still needed. Herein, we studied cationic conjugated polymer (CCP)-based binding assays taking advantage of the conformational change of aptamer after binding with a target, which is reflected by the fluorescence change of the CCP. Poly(3-(3′-N,N,N-triethylamino-1′-propyloxy)-4-methyl-2,5-thiophene hydrochloride) (PMNT) was used as a model CCP in this study, and the optimal buffer was close to physiological conditions with 100 mM NaCl and 10 mM MgCl2. We characterized four aptamers for K+, adenosine, cortisol, and caffeine. For cortisol and caffeine, the drop in the 580 nm peak intensity was used for quantification, whereas for K+ and adenosine, the fluorescence ratio at 580 over 530 nm was used. The longer stem of the stem-loop structured aptamer facilitated binding of the target and enlarged the detection signal. High specificity was achieved in differentiating targets with analogues. Compared with the SYBR Green I dye-based staining method, our method achieved equal or even higher sensitivity. Therefore, this assay is practicable as a general aptamer binding assay. The simple, label-free, quick response, and cost-effective features will make it a useful method to evaluate aptamer binding. At the same time, this system can also serve as label-free biosensors for target detection.
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