More DNA-Aptamers for Small Drugs: A Capture-SELEX Coupled with Surface Plasmon Resonance and High-Throughput Sequencing.
ACS Comb Sci
; 17(5): 326-33, 2015 May 11.
Article
en En
| MEDLINE
| ID: mdl-25875077
ABSTRACT
To address limitations in the production of DNA aptamers against small molecules, we introduce a DNA-based capture-SELEX (systematic evolution of ligands by exponential enrichment) protocol with long and continuous randomized library for more flexibility, coupled with in-stream direct-specificity monitoring via SPR and high throughput sequencing (HTS). Applying this capture-SELEX on tobramycin shows that target-specificity arises at cycle number 8, which is confirmed by sequence convergence in HTS analysis. Interestingly, HTS also shows that the most enriched sequences are already visible after only two capture-SELEX cycles. The best aptamers displayed K(D) of approximately 200 nM, similar to RNA and DNA-based aptamers previously selected for tobramycin. The lowest concentration of tobramycin detected on label-free SPR experiments with the selected aptamers is 20-fold smaller than the clinical range limit, demonstrating suitability for small-drug biosensing.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
ADN
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Resonancia por Plasmón de Superficie
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Aptámeros de Nucleótidos
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Técnica SELEX de Producción de Aptámeros
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Ensayos Analíticos de Alto Rendimiento
Tipo de estudio:
Clinical_trials
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Guideline
Idioma:
En
Revista:
ACS Comb Sci
Año:
2015
Tipo del documento:
Article
País de afiliación:
Suiza