Three different signal amplification strategies for the impedimetric sandwich detection of thrombin
Ocaña Tejada, Cristina (Universitat Autònoma de Barcelona. Departament de Química)
Valle Zafra, Manuel del (Universitat Autònoma de Barcelona. Departament de Química)

Data:	2016
Resum:	In this work, we report a comparative study on three highly specific amplification strategies for the ultrasensitive detection of thrombin with the use of aptamer sandwich protocol. The protocol consisted on the use of a first thrombin aptamer immobilized on the electrode surface, the recognition of thrombin protein, and the reaction with a second biotinylated thrombin aptamer forming the sandwich. Through the exposed biotin end, three variants have been tested to amplify the electrochemical impedance signal. The strategies included (a) silver enhancement treatment, (b) gold enhancement treatment and (c) insoluble product produced by the combination of the enzyme horseradish peroxidase (HRP) and 3-amino-9-ethylcarbazole (AEC). The properties of the sensing surface were probed by electrochemical impedance measurements in the presence of the ferrocyanide/ferricyanide redox marker. Insoluble product strategy and silver enhancement treatment resulted in the lowest detection limit (0. 3 pM), while gold enhancement method resulted in the highest reproducibility, 8. 8% RSD at the pM thrombin concentration levels. Results of silver and gold enhancement treatment also permitted direct inspection by scanning electron microscopy (SEM).
Ajuts:	Ministerio de Economía y Competitividad CTQ2013-41577-P
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial i la comunicació pública de l'obra, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. No es permet la creació d'obres derivades.
Llengua:	Anglès
Document:	Article ; recerca ; Versió acceptada per publicar
Matèria:	Aptamer ; Impedance ; Gold nanoparticles ; Amplification ; Aptasensor ; Insoluble product
Publicat a:	Analytica chimica acta, Vol. 912 (March 2016) , p. 117-124, ISSN 1873-4324

DOI: 10.1016/j.aca.2016.01.027

22 p, 3.1 MB

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