Aptamer based vanillin sensor using an ion-sensitive field-effect transistor

Aptamer based vanillin sensor using an ion-sensitive field-effect transistor An aptamer for vanillin was obtained and then used for the development of an aptasensor based on an ion-sensitive field-effect transistor (ISFET). This aptamer (a single-stranded DNA;ssDNA) was selected using the Capture-SELEX protocol, which suites well for selection of aptamers to small molecules. Among six aptamer candidates, the aptamer Van_74 with the highest affinity for vanillin was chosen (elution of 35% of the aptamer from a solid support in the presence of 2 mM of vanillin). Van_74 was characterized using nondenaturating PAGE of washouts from magnetic beads. It is shown that Van_74 binds to vanillin with an dissociation constant of >7.8 μM (determined by nondenaturating PAGE) and it was specific to vanillin in comparison with interferents: benzaldehyde, guaiacol, furaneol, ethyl guaiacol and ethyl vanillin. Also it was shown that change of buffer composition greatly affected the binding ability of Van_74. For biosensor fabrication aptamer was immobilised on the Ta2O5-sensitive surface of the ISFET via “click-chemistry”. Detection scheme implied dehybridisation of the ssDNA probe from the aptamer and release in the solution during the addition of vanillin. As a result, the surface potential increase upon vanillin binding with the aptamer was detected by the transistor. The biosensor had a detection limit of 1.55 × 10−7 M and a dynamic range from 1.55 × 10−7 M to 1 × 10−6 M. Effective constant Kd,eff for vanillin binding on biosensor surface was calculated to be (9 ± 3) × 10−7 M. This allows selective detection of vanillin in the mixture of interferents and in samples of coffee extract. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Microchimica Acta Springer Journals

Aptamer based vanillin sensor using an ion-sensitive field-effect transistor

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Publisher
Springer Vienna
Copyright
Copyright © 2017 by Springer-Verlag GmbH Austria, part of Springer Nature
Subject
Chemistry; Nanochemistry; Nanotechnology; Characterization and Evaluation of Materials; Analytical Chemistry; Microengineering
ISSN
0026-3672
eISSN
1436-5073
D.O.I.
10.1007/s00604-017-2586-4
Publisher site
See Article on Publisher Site

Abstract

An aptamer for vanillin was obtained and then used for the development of an aptasensor based on an ion-sensitive field-effect transistor (ISFET). This aptamer (a single-stranded DNA;ssDNA) was selected using the Capture-SELEX protocol, which suites well for selection of aptamers to small molecules. Among six aptamer candidates, the aptamer Van_74 with the highest affinity for vanillin was chosen (elution of 35% of the aptamer from a solid support in the presence of 2 mM of vanillin). Van_74 was characterized using nondenaturating PAGE of washouts from magnetic beads. It is shown that Van_74 binds to vanillin with an dissociation constant of >7.8 μM (determined by nondenaturating PAGE) and it was specific to vanillin in comparison with interferents: benzaldehyde, guaiacol, furaneol, ethyl guaiacol and ethyl vanillin. Also it was shown that change of buffer composition greatly affected the binding ability of Van_74. For biosensor fabrication aptamer was immobilised on the Ta2O5-sensitive surface of the ISFET via “click-chemistry”. Detection scheme implied dehybridisation of the ssDNA probe from the aptamer and release in the solution during the addition of vanillin. As a result, the surface potential increase upon vanillin binding with the aptamer was detected by the transistor. The biosensor had a detection limit of 1.55 × 10−7 M and a dynamic range from 1.55 × 10−7 M to 1 × 10−6 M. Effective constant Kd,eff for vanillin binding on biosensor surface was calculated to be (9 ± 3) × 10−7 M. This allows selective detection of vanillin in the mixture of interferents and in samples of coffee extract.

Journal

Microchimica ActaSpringer Journals

Published: Dec 2, 2017

References

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