1. Monovalent labeling system improves the sensitivity of aptamer-based inhibition assays for small molecule detection.
- Author
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González-Fernández, Eva, de-los-Santos-Álvarez, Noemí, Miranda-Ordieres, Arturo José, and Lobo-Castañón, María Jesús
- Subjects
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APTAMERS , *BIOLOGICAL assay , *MOLECULES , *ELECTROCHEMICAL sensors , *TOBRAMYCIN , *BIOCONJUGATES , *ENZYME analysis - Abstract
Abstract: 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 5μM to 0.1μM 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. [Copyright &y& Elsevier]
- Published
- 2013
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