2024-03-04 ノースカロライナ州立大学(NCState)
<関連情報>
- https://news.ncsu.edu/2024/03/dna-aptamer-drug-sensors-can-instantly-detect-cocaine-heroin-and-fentanyl-even-when-combined-with-other-drugs/
- https://pubs.acs.org/doi/full/10.1021/jacsau.3c00801
- https://pubs.acs.org/doi/abs/10.1021/jacs.3c11350
アプタマーベースの比色オピオイド検査法の開発 Developing Aptamer-Based Colorimetric Opioid Tests
Juan Canoura, Obtin Alkhamis, Matthew Venzke, Phuong T. Ly, and Yi Xiao
JACS Au Published:March 1, 2024
DOI:https://doi.org/10.1021/jacsau.3c00801
Abstract
Opioids collectively cause over 80,000 deaths in the United States annually. The ability to rapidly identify these compounds in seized drug samples on-site will be essential for curtailing trafficking and distribution. Chemical reagent-based tests are fast and simple but also notorious for giving false results due to poor specificity, whereas portable Raman spectrometers have excellent selectivity but often face interference challenges with impure drug samples. In this work, we develop on-site sensors for morphine and structurally related opioid compounds based on in vitro-selected oligonucleotide affinity reagents known as aptamers. We employ a parallel-and-serial selection strategy to isolate aptamers that recognize heroin, morphine, codeine, hydrocodone, and hydromorphone, along with a toggle-selection approach to isolate aptamers that bind oxycodone and oxymorphone. We then utilize a new high-throughput sequencing-based approach to examine aptamer growth patterns over the course of selection and a high-throughput exonuclease-based screening assay to identify optimal aptamer candidates. Finally, we use two high-performance aptamers with KD of ∼1 μM to develop colorimetric dye-displacement assays that can specifically detect opioids like heroin and oxycodone at concentrations as low as 0.5 μM with a linear range of 0–16 μM. Importantly, our assays can detect opioids in complex chemical matrices, including pharmaceutical tablets and drug mixtures; in contrast, the conventional Marquis test completely fails in this context. These aptamer-based colorimetric assays enable the naked-eye identification of specific opioids within seconds and will play an important role in combatting opioid abuse.
コカインを生体内および生体外で感知する高親和性アプタマー High-Affinity Aptamers for In Vitro and In Vivo Cocaine Sensing
Obtin Alkhamis, Juan Canoura, Yuyang Wu, Nicole A. Emmons, Yuting Wang, Kevin M. Honeywell, Kevin W. Plaxco, Tod E. Kippin, and Yi Xiao
Journal of the American Chemical Society Published:January 26, 2024
DOI:https://doi.org/10.1021/jacs.3c11350
Abstract
The ability to quantify cocaine in biological fluids is crucial for both the diagnosis of intoxication and overdose in the clinic as well as investigation of the drug’s pharmacological and toxicological effects in the laboratory. To this end, we have performed high-stringency in vitro selection to generate DNA aptamers that bind cocaine with nanomolar affinity and clinically relevant specificity, thus representing a dramatic improvement over the current-generation, micromolar-affinity, low-specificity cocaine aptamers. Using these novel aptamers, we then developed two sensors for cocaine detection. The first, an in vitro fluorescent sensor, successfully detects cocaine at clinically relevant levels in 50% human serum without responding significantly to other drugs of abuse, endogenous substances, or a diverse range of therapeutic agents. The second, an electrochemical aptamer-based sensor, supports the real-time, seconds-resolved measurement of cocaine concentrations in vivo in the circulation of live animals. We believe the aptamers and sensors developed here could prove valuable for both point-of-care and on-site clinical cocaine detection as well as fundamental studies of cocaine neuropharmacology.
