Solid-contact potentiometric aptasensor based on aptamer functionalized carbon nanotubes for the direct determination of proteins.

A facile, solid-contact selective potentiometric aptasensor exploiting a network of single-walled carbon nanotubes (SWCNT) acting as a transducing element is described in this work. The molecular properties of the SWCNT surface have been modified by covalently linking aptamers as biorecognition elements to the carboxylic groups of the SWCNT walls. As a model system to demonstrate the generic application of the approach, a 15-mer thrombin aptamer interacts with thrombin and the affinity interaction gives rise to a direct potentiometric signal that can be easily recorded within 15 s. The dynamic linear range, with a sensitivity of 8.0 mV/log a(Thr) corresponds to the 10(-7)-10(-6) M range of thrombin concentrations, with a limit of detection of 80 nM. The aptasensor displays selectivity against elastase and bovine serum albumin and is easily regenerated by immersion in 2 M NaCl. The aptasensor demonstrates the capacity of direct detection of the recognition event avoiding the use of labels, mediators, or the addition of further reagents or analyte accumulation.

[1]  Ronghua Yang,et al.  Noncovalent assembly of carbon nanotubes and single-stranded DNA: an effective sensing platform for probing biomolecular interactions. , 2008, Analytical chemistry.

[2]  K. Yasuda,et al.  Evaluation of the number of carboxyl groups on glassy carbon after modification by 3,4-dihydroxybenzylamine , 2007 .

[3]  Cees Dekker,et al.  Nanotechnology: Carbon nanotubes with DNA recognition , 2002, Nature.

[4]  Kenzo Maehashi,et al.  Label-free protein biosensor based on aptamer-modified carbon nanotube field-effect transistors. , 2007, Analytical chemistry.

[5]  Cees Dekker,et al.  Identifying the mechanism of biosensing with carbon nanotube transistors. , 2008, Nano letters.

[6]  Jeong-O Lee,et al.  Single-walled carbon nanotube biosensors using aptamers as molecular recognition elements. , 2005, Journal of the American Chemical Society.

[7]  F. Rius,et al.  Transduction mechanism of carbon nanotubes in solid-contact ion-selective electrodes. , 2009, Analytical chemistry.

[8]  Ernö Pretsch,et al.  Aptamer-based potentiometric measurements of proteins using ion-selective microelectrodes. , 2008, Analytical chemistry.

[9]  P. Mair,et al.  Plasma elastase concentrations and pulmonary function after cardiopulmonary bypass. , 1994, The Journal of thoracic and cardiovascular surgery.

[10]  D. Walt,et al.  A fiber-optic microarray biosensor using aptamers as receptors. , 2000, Analytical biochemistry.

[11]  M. Klein,et al.  Probing the structure of DNA-carbon nanotube hybrids with molecular dynamics. , 2007, Nano letters.

[12]  Grégoire Herzog,et al.  Electrochemical strategies for the label-free detection of amino acids, peptides and proteins. , 2007, The Analyst.

[13]  Róbert E. Gyurcsányi,et al.  Quality control criteria for solid-contact, solvent polymeric membrane ion-selective electrodes , 2009 .

[14]  D. Tang,et al.  Direct and Rapid Detection of Diphtherotoxin via Potentiometric Immunosensor Based on Nanoparticles Mixture and Polyvinyl Butyral as Matrixes , 2005 .

[15]  Ciara K O'Sullivan,et al.  Reusable impedimetric aptasensor. , 2005, Analytical chemistry.

[16]  E. Pretsch The new wave of ion-selective electrodes , 2007 .

[17]  P. Eklund,et al.  Debundling and dissolution of single-walled carbon nanotubes in amide solvents. , 2004, Journal of the American Chemical Society.

[18]  F. Rius,et al.  Ion-selective electrodes using carbon nanotubes as ion-to-electron transducers. , 2008, Analytical chemistry.

[19]  Yanxiu Zhou,et al.  A potentiometric protein sensor built with surface molecular imprinting method. , 2008, Biosensors & bioelectronics.

[20]  T. Hianik,et al.  Influence of ionic strength, pH and aptamer configuration for binding affinity to thrombin. , 2007, Bioelectrochemistry.

[21]  A. Tulinsky,et al.  The structure of alpha-thrombin inhibited by a 15-mer single-stranded DNA aptamer. , 1994, The Journal of biological chemistry.

[22]  M. Mascini,et al.  Aptamers-based assays for diagnostics, environmental and food analysis. , 2007, Biomolecular engineering.

[23]  A. Guiseppi-Elie,et al.  Potentiometric monitoring DNA hybridization. , 2009, Biosensors & bioelectronics.

[24]  M. Zheng,et al.  DNA-assisted dispersion and separation of carbon nanotubes , 2003, Nature materials.

[25]  H. Gunasingham,et al.  Comparative study of glassy carbon as an electrode material , 1982 .