Efficient biorecognition of thrombin (TB), a serine protease with crucial role in physiological and pathological blood coagulation, is a hot topic in medical diagnostics. In this work, we investigate the ability of synthetic thrombin aptamer (TBA), immobilized on a gold substrate, to bind thrombin by two different label-free techniques: the quartz crystal microbalance (QCM) and the spectroscopic ellipsometry (SE). By QCM characterization in the range from 20 to 110 nM, we demonstrate high specificity of TBA-TB interaction and determine affinity constant (Kd) of 17.7 ± 0.3 nM, system sensitivity of 0.42 ± 0.03 Hz nM-1, and limit of detection (LOD) of 240 ± 20 pM. The interaction between TBA and TB is also investigated by SE, an all-optical method, by quantifying the thickness increase of the TBA film assembled on gold substrate. AFM characterization of TBA and TB molecules deposited on flat silicon surface is also supplied.

Nanogravimetric and Optical Characterizations of Thrombin Interaction with a Self-Assembled Thiolated Aptamer / Politi, Jane; Rea, Ilaria; Nici, Fabrizia; Dardano, Principia; Terracciano, Monica; Oliviero, Giorgia; Borbone, Nicola; Piccialli, Gennaro; DE STEFANO, Luca. - In: JOURNAL OF SENSORS. - ISSN 1687-725X. - 2016:(2016), pp. 1-8. [10.1155/2016/3561863]

Nanogravimetric and Optical Characterizations of Thrombin Interaction with a Self-Assembled Thiolated Aptamer

REA, ILARIA;NICI, FABRIZIA;DARDANO, PRINCIPIA;TERRACCIANO, MONICA;OLIVIERO, GIORGIA;BORBONE, NICOLA;PICCIALLI, GENNARO;DE STEFANO, Luca
2016

Abstract

Efficient biorecognition of thrombin (TB), a serine protease with crucial role in physiological and pathological blood coagulation, is a hot topic in medical diagnostics. In this work, we investigate the ability of synthetic thrombin aptamer (TBA), immobilized on a gold substrate, to bind thrombin by two different label-free techniques: the quartz crystal microbalance (QCM) and the spectroscopic ellipsometry (SE). By QCM characterization in the range from 20 to 110 nM, we demonstrate high specificity of TBA-TB interaction and determine affinity constant (Kd) of 17.7 ± 0.3 nM, system sensitivity of 0.42 ± 0.03 Hz nM-1, and limit of detection (LOD) of 240 ± 20 pM. The interaction between TBA and TB is also investigated by SE, an all-optical method, by quantifying the thickness increase of the TBA film assembled on gold substrate. AFM characterization of TBA and TB molecules deposited on flat silicon surface is also supplied.
2016
Nanogravimetric and Optical Characterizations of Thrombin Interaction with a Self-Assembled Thiolated Aptamer / Politi, Jane; Rea, Ilaria; Nici, Fabrizia; Dardano, Principia; Terracciano, Monica; Oliviero, Giorgia; Borbone, Nicola; Piccialli, Gennaro; DE STEFANO, Luca. - In: JOURNAL OF SENSORS. - ISSN 1687-725X. - 2016:(2016), pp. 1-8. [10.1155/2016/3561863]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11588/672680
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