Toggled RNA Aptamers Against Aminoglycosides Allowing Facile Detection of Antibiotics Using Gold Nanoparticle Assays

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• 作者：Nicola Derbyshire ; Simon J. White ; David H. J. Bunka ; Lei Song ; Sara Stead ; Jonathan Tarbin ; Matthew Sharman ; Dejian Zhou ; Peter G. Stockley
• 刊名：Analytical Chemistry
• 出版年：2012
• 出版时间：August 7, 2012
• 年：2012
• 卷：84
• 期：15
• 页码：6595-6602
• 全文大小：407K
• 年卷期：v.84,no.15(August 7, 2012)
• ISSN：1520-6882

文摘

We have used systematic evolution of ligands by exponential enrichment (SELEX) to isolate RNA aptamers against aminoglycoside antibiotics. The SELEX rounds were toggled against four pairs of aminoglycosides with the goal of isolating reagents that recognize conserved structural features. The resulting aptamers bind both of their selection targets with nanomolar affinities. They also bind the less structurally related targets, although they show clear specificity for this class of antibiotics. We show that this lack of aminoglycoside specificity is a common property of aptamers previously selected against single compounds and described as 鈥渟pecific鈥? Broad target specificity aptamers would be ideal for sensors detecting the entire class of aminoglycosides. We have used ligand-induced aggregation of gold-nanoparticles coated with our aptamers as a rapid and sensitive assay for these compounds. In contrast to DNA aptamers, unmodified RNA aptamers cannot be used as the recognition ligand in this assay, whereas 2鈥?fluoro-pyrimidine derivatives work reliably. We discuss the possible application of these reagents as sensors for drug residues and the challenges for understanding the structural basis of aminoglycoside-aptamer recognition highlighted by the SELEX results.