Monovalent labeling system improves the sensitivity of aptamer-based inhibition assays for small molecule detection

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• 作者：Eva Gonzá ; lez-Ferná ; ndez ^{gfernandezeva@hotmail.com}Author Vitae ; Noemí ; de-los-Santos-Á ; lvarez ^{santosnoemi@uniovi.es}Author Vitae ; Arturo José ; Miranda-Ordieres ^{amir@uniovi.es}Author Vitae ; Marí ; a Jesú ; s Lobo-Castañ ; ó ; n ; ^{mjlc@uniovi.es} ; ^{mjloboc@gmail.com}Author Vitae
• 关键词：Aptamer ; Electrochemical detection ; Magnetic particles ; Monovalent labeling ; Small molecule ; Tobramycin
• 刊名：Sensors and Actuators B: Chemical
• 出版年：2013
• 出版时间：June, 2013
• 年：2013
• 卷：182
• 期：Complete
• 页码：668-674
• 全文大小：586 K

文摘

The design and performance of an inhibition assay for the electrochemical detection of tobramycin is reported. This platform uses a monovalent system for introducing the enzyme conjugate on a tagged-aptamer that specifically recognizes the aminoglycoside antibiotic tobramycin. Compared with multivalent systems such as biotin-streptavidin, the sensitivity is greatly improved reducing the limit of detection from 5 ¦ÌM to 0.1 ¦ÌM in aqueous solution. This labeling strategy along with the use of a nuclease-resistant RNA aptamer allowed the determination of tobramycin in pre-treated human serum samples within the therapeutic range, using magnetic microparticles as a solid support for the aptassay, and differential pulse voltammetry as detection technique.