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Chemosensors 2016, 4(3), 18; doi:10.3390/chemosensors4030018

Miniaturized Aptamer-Based Assays for Protein Detection

1
Elettra-Sincrotone S.C.p.A., SS 14 km 163,5 in AREA Science Park, Basovizza, Trieste 34149, Italy
2
Department of Physics, University of Trieste, Via Valerio 9, Trieste 34127, Italy
3
INSTM-ST Unit, SS 14 km 163,5 in AREA Science Park, Basovizza, Trieste 34149, Italy
4
Centre of Excellence for Biosensors, Instrumentation and Process Control, Tovarniška 26, Ajdovščina SI-5270, Slovenia
*
Author to whom correspondence should be addressed.
Academic Editors: Paolo Ugo and Ligia Moretto
Received: 11 July 2016 / Revised: 12 August 2016 / Accepted: 29 August 2016 / Published: 2 September 2016
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)
View Full-Text   |   Download PDF [3255 KB, uploaded 2 September 2016]   |  

Abstract

The availability of devices for cancer biomarker detection at early stages of the disease is one of the most critical issues in biomedicine. Towards this goal, to increase the assay sensitivity, device miniaturization strategies empowered by the employment of high affinity protein binders constitute a valuable approach. In this work we propose two different surface-based miniaturized platforms for biomarker detection in body fluids: the first platform is an atomic force microscopy (AFM)-based nanoarray, where AFM is used to generate functional nanoscale areas and to detect biorecognition through careful topographic measurements; the second platform consists of a miniaturized electrochemical cell to detect biomarkers through electrochemical impedance spectroscopy (EIS) analysis. Both devices rely on robust and highly-specific protein binders as aptamers, and were tested for thrombin detection. An active layer of DNA-aptamer conjugates was immobilized via DNA directed immobilization on complementary single-stranded DNA self-assembled monolayers confined on a nano/micro area of a gold surface. Results obtained with these devices were compared with the output of surface plasmon resonance (SPR) assays used as reference. We succeeded in capturing antigens in concentrations as low as a few nM. We put forward ideas to push the sensitivity further to the pM range, assuring low biosample volume (μL range) assay conditions. View Full-Text
Keywords: biosensors; aptamers; AFM; nanoarray; EIS biosensors; aptamers; AFM; nanoarray; EIS
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Bosco, A.; Ambrosetti, E.; Mavri, J.; Capaldo, P.; Casalis, L. Miniaturized Aptamer-Based Assays for Protein Detection. Chemosensors 2016, 4, 18.

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