期刊
ANALYTICAL CHEMISTRY
卷 93, 期 6, 页码 3172-3180出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.0c04592
关键词
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资金
- National Institute of Justice, Grant Office of Justice Programs, U.S. Department of Justice [2015-R2-CX-0034]
- National Institutes of Health, National Institute on Drug Abuse [R21DA04533401A1]
Aptamers, isolated through the SELEX process, exhibit high affinity and specificity for challenging small-molecule targets. Natural DNA libraries can yield high-quality aptamers, demonstrating an effective isolation workflow.
Aptamers are nucleic acid-based affinity reagents that are isolated via an in vitro process known as systematic evolution of ligands by exponential enrichment (SELEX). Despite their great potential for a wide range of analytical applications, there are relatively few high-quality small-molecule binding aptamers, especially for challenging targets that have low water solubility and/or limited moieties for aptamer recognition. The use of libraries containing chemically modified bases may improve the outcome of some SELEX experiments, but this approach is costly and yields inconsistent results. Here, we demonstrate that a thoughtfully designed SELEX procedure with natural DNA libraries can isolate aptamers with high affinity and specificity for challenging small molecules, including targets for which such selections have previously failed. We first isolate a DNA aptamer with nanomolar affinity and high specificity for (-)-trans-Delta(9) -tetrahydrocannabinol (THC), a target previously thought to be unsuitable for SELEX with natural DNA libraries. We subsequently isolate aptamers that exhibit high affinity and cross-reactivity to two other challenging targets, synthetic cannabinoids UR-144 and XLR-11, while maintaining excellent specificity against a wide range of non-target interferents. Our findings demonstrate that natural nucleic acid libraries can yield high-quality aptamers for small-molecule targets, and we outline a robust workflow for isolating other such aptamers in future selection efforts.
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