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Investigation of the selectivity of thrombin-binding aptamers for thrombin titration in murine plasma

Authors :
Ana Trapaidze
Jean-Pascal Herault
Anne Marie Gue
Jean-Marc Herbert
Aurélien Bancaud
Équipe Nano Ingénierie et Intégration des Systèmes (LAAS-N2IS)
Laboratoire d'analyse et d'architecture des systèmes (LAAS)
Université Toulouse Capitole (UT Capitole)
Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse)
Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université Toulouse - Jean Jaurès (UT2J)
Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3)
Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP)
Université de Toulouse (UT)-Université Toulouse Capitole (UT Capitole)
Université de Toulouse (UT)
SANOFI Recherche
Équipe Micro-Nanofluidique pour les sciences de la vie et de l’environnement (LAAS-MILE)
Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse 1 Capitole (UT1)
Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3)
Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse)
Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National Polytechnique (Toulouse) (Toulouse INP)
Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse 1 Capitole (UT1)
Université Fédérale Toulouse Midi-Pyrénées
Source :
Biosensors and Bioelectronics, Biosensors and Bioelectronics, 2016, 78, pp.58-66. ⟨10.1016/j.bios.2015.11.017⟩, Biosensors and Bioelectronics, Elsevier, 2016, 78, pp.58-66. ⟨10.1016/j.bios.2015.11.017⟩
Publication Year :
2016
Publisher :
Elsevier BV, 2016.

Abstract

International audience; Detection of thrombin in plasma raises timely challenges to enable therapeutic management of thrombosis in patients under vital threat. Thrombin binding aptamers represent promising candidates as sensing elements for the development of real-time thrombin biosensors; however implementation of such biosensor requires the clear understanding of thrombin-aptamer interaction properties in real-like environment. In this study, we used Surface Plasmon Resonance technique to answer the questions of specificity and sensitivity of thrombin detection by the thrombin-binding aptamers HD1, NU172 and HD22. We systematically characterized their properties in the presence of thrombin, as well as interfering molecular species such as the thrombin precursor prothrombin, thrombin in complex with some of its natural inhibitors, nonspecific serum proteins, and diluted plasma. Kinetic experiments show the multiple binding modes of HD1 and NU172, which both interact with multiple sites of thrombin with low nanomolar affinities and show little specificity of interaction for prothrombin vs. thrombin. HD22, on the other hand, binds specifically to thrombin exosite II and has no affinity to prothrombin at all. While thrombin in complex with some of its inhibitors could not be recognized by any aptamer, the binding of HD1 and NU172 properties is compromised by thrombin inhibitors alone, as well as with serum albumin. Finally, the complex nature of plasma was overwhelming for HD1, but we define conditions for the thrombin detection at 10nM range in 100-fold diluted plasma by HD22. Consequently HD22 showed key advantage over HD1 and NU172, and appears as the only alternative to design an aptasensor.

Details

ISSN :
09565663
Volume :
78
Database :
OpenAIRE
Journal :
Biosensors and Bioelectronics
Accession number :
edsair.doi.dedup.....a5cf8312b3bef439ddde564a75416f14
Full Text :
https://doi.org/10.1016/j.bios.2015.11.017