Aptamer adaptive binding assessed by stilbene photoisomerization towards regenerating aptasensors

SENSORS AND ACTUATORS B-CHEMICAL

卷:257

Article

Zhou, Yubin¹  Wu, Yuanyuan¹  Pokholenko, Oleksandr¹  Grimsrud, Marissa²  Sham, Yuk^2,3  Papper, Vladislav¹  Marks, Robert⁴  Steele, Terry¹ 

[1] Nanyang Technol Univ, Coll Engn, Sch Mat Sci & Engn, 50 Nanyang Ave, Singapore 639798, Singapore

[2] Univ Minnesota, Bioinformat & Computat Biol Program, Minneapolis, MN 55455 USA

[3] Univ Minnesota, Med Sch, Dept Integrat Biol & Physiol, 2231 6th St SE, Minneapolis, MN 55455 USA

[4] Ben Gurion Univ Negev, Fac Engn Sci, Dept Biotechnol Engn, POB 653, IL-84105 Beer Sheva, Israel

关键词: Aptamer;    Stilbene;    Fluorescence decay;    Photoisomerization;    Aptasensor;    Malachite green;   

DOI  :  10.1016/j.snb.2017.10.135

来源: Elsevier

【 摘 要 】

Fluorescent aptasensors are reliant on static fluorescence intensity measurements, which suffer drawbacks such as background interference and laborious separation procedures. A unique aptasensor based on the photochrome aptamer switch assay (PHASA) has been developed that is independent of background fluorescence, requires no analyte separation, and allows rapid quantification within seconds. Malachite green aptamer (MGA) conjugated with a water-soluble stilbene on the MGA 3' C38 terminus was chosen for building the proof-of-concept aptasensor. In the presence of malachite green and tetramethylrosamine ligands, the rate of the stilbene fluorescence decay was found to be linearly dependent on the ligand concentration. Molecular dynamic simulation suggests hydrogen bonding between stilbene sulfonates and neighboring nucleotides is the primary mechanism responsible for rate changes in stilbene photoisomerization. Analysis of the apparent fluorescence decay rate (k(app)) versus analyte concentration gives a limit of detection (LOD) of 2 mu M for MG and 0.6 mu M for TMR. This aptasensor design opens up a new sensing mode, which is promising for rapid development of SELEX generated molecular recognition elements. (C) 2017 Elsevier B.V. All rights reserved.

【 授权许可】

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