Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/143105
Title: Analysis of the role played by ligand-induced folding of the cocaine-binding aptamer in the photochrome aptamer switch assay
Authors: Shoara, Aron A.
Churcher, Zachary R.
Steele, Terry W. J.
Johnson, Philip E.
Keywords: Engineering::Materials
Issue Date: 2020
Source: Shoara, A. A., Churcher, Z. R., Steele, T. W. J., & Johnson, P. E. (2020). Analysis of the role played by ligand-induced folding of the cocaine-binding aptamer in the photochrome aptamer switch assay. Talanta, 217, 121022. doi:10.1016/j.talanta.2020.121022
Journal: Talanta
Abstract: The Photochrome Aptamer Switch Assay (PHASA) relies on ligand binding by an aptamer to alter the local environment of a stilbene compound covalently attached to the 5' end of the aptamer. We used the PHASA with both structure switching and non-structure switching versions of the cocaine-binding aptamer. We show that the largest change in fluorescence intensity and the lowest concentration limit of detection (CLooD) is obtained using the structure-switching cocaine-binding aptamer. Fluorescence anisotropy measurements were used to quantify the affinity of the conjugated aptamer to cocaine. We also used thermal melt analysis and Nuclear Magnetic Resonance (NMR) spectroscopy to show that the addition of the stilbene to the aptamer increases the melt temperature of the cocaine-bound structure-switching aptamer by (6.4 ± 0.3) °C compared to the unconjugated aptamer while the free form of the structure-switching aptamer-stilbene conjugate remains unfolded.
URI: https://hdl.handle.net/10356/143105
ISSN: 1873-3573
DOI: 10.1016/j.talanta.2020.121022
Rights: © 2020 Elsevier. All rights reserved. This paper was published in Talanta and is made available with permission of Elsevier.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:MSE Journal Articles

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