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Nanostructured Platform Based on Graphene-polypyrrole Composite for Immunosensor Fabrication
Available online at www.sciencedirect.com
ScienceDirect Procedia Technology 27 (2017) 108 – 109
Biosensors 2016
Nanostructured platform based on graphene-polypyrrole composite for immunosensor fabrication Andreea Cernat, Mihaela Tertiș, Claudia Nicoleta Păpară, Ede Bodoki, Robert Săndulescu* Analytical Chemistry Department, "Iuliu Haţieganu" University of Medicine and Pharmacy, 4 Pasteur Street, 400349, Cluj-Napoca, Romania
Abstract A hybrid graphene-polypyrrole (PPy) composite-based platform was elaborated by using nanosphere lithography structuration. This platform was further used for the antibody anti-acetaminophen immobilization in order to obtain an immunosensor for selective detection of paracetamol (acetaminophen) by using electrochemical methods. After patterning of the composite layer, the five-fold improvement of electrochemical signal was observed, suggesting that a higher amount of antibody was immobilized. This strategy permitted the ease and controlled immobilization of bioreceptors on the composite graphene-PPy platform and enhanced the sensitivity for paracetamol detection. © 2017 2016The TheAuthors. Authors.Published Published Elsevier © byby Elsevier Ltd.Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Peer-review under responsibility of the organizing committee of Biosensors 2016. Peer-review under responsibility of the organizing committee of Biosensors 2016 Keywords: graphene; polypyrrole; immunosensor; acetaminophen; nanosphere lithography
The entrapment of functionalized graphene in polypyrrole (PPy) conductive layers improves their engineering features. The association of graphene-PPy composite with nanosphere lithography generated a hybrid polystyrenegraphene-PPy patterned platform, with a further enhancement of their characteristics: an improved mechanical stability and electron transfer rate, combined with an increase of the active surface area >1, 2@. The immobilization of activated graphene oxide sheets onto the electrode was performed through electrostatic bonds, while the anti-paracetamol antibody (ApAb) biocomponent molecules were covalently bounded after the substrate modification with N-hydroxysuccinimide activated carboxylic functional groups. This strategy permits the controlled immobilization of ApAb in order to elaborate an immunosensor for paracetamol detection.
* Corresponding author. Tel.: +40 264 593118; fax: +40 264 597257. E-mail address: [email protected]
2212-0173 © 2017 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Peer-review under responsibility of the organizing committee of Biosensors 2016 doi:10.1016/j.protcy.2017.04.047
109
Andreea Cernat et al. / Procedia Technology 27 (2017) 108 – 109
The behavior of the nanostructured hybrid platform was tested by imagistic (Figure 1 B), impedimetric, and electrochemical quartz crystal microbalance experiments proving the successful patterning of the composite material and the biomolecule’s immobilization.
A
C B Fig. 1. (A) The development of the nanostructured platform and the immunosensor fabrication; (B) Optical microscopy images performed on the PS-graphene-PPy composite and (C) Current intensity variation, following the incubation with 5 µM paracetamol solution, recorded in PBS at the unstructured graphene-PPy (black line) and the nanostructured graphene-PPy (red line); (ApAb was incubated for 60 minutes; 5 % BSA for 20 minutes and paracetamol for 15 minutes) [3]
The unstructured and nanostructured graphene-PPy hybrid platforms were elaborated and the properties were tested by using paracetamol, a stable molecule with electrochemical activity. All the steps involved in immunosensor elaboration, presented in Figure 1 A, were optimized and it was observed that the sensor allowed the specific detection of paracetamol in a linear range of 0.1 to 25 µM at the unstructured platform. The analytical signal was of about 5 times improved on the patterned material in comparison with the unstructured one, proving the superior features of the structured surface (Figure 1 C). This nanopatterning strategy and the possibility of ease and controlled immobilization of bioreceptors may be exploited in biosensors development and applied in biomedical domain for the detection of biomarkers. Acknowledgements The authors are grateful for the financial support from the Romanian National Authority for Scientific Research, for project number PN-II-ID-PCE-2011-3-0355.
References [1] Cernat A, Griveau S, Martin P, Lacroix JC, Farcău C, Săndulescu R, Bedioui F. Electrografted nanostructured platforms for click chemistry. Electrochem Commun 2012;23:141-4. [2] Cernat A, Le Goff A, Holzinger M, Săndulescu R, Cosnier S. Micro-to nanostructured poly(pyrrole-nitrilotriacetic acid) films via nanosphere templates: applications to 3D enzyme attachment by affinity interactions. Anal Bioanal Chem 2014;406:1141-7. [3] Cernat A, Tertiş M, Păpară CN, Bodoki E, Săndulescu R. Nanostructured platform based on graphene-polypyrrole composite for immunosensor fabrication. Int J Electrochem Sci 2015;10:4718-31.
ScienceDirect Procedia Technology 27 (2017) 108 – 109
Biosensors 2016
Nanostructured platform based on graphene-polypyrrole composite for immunosensor fabrication Andreea Cernat, Mihaela Tertiș, Claudia Nicoleta Păpară, Ede Bodoki, Robert Săndulescu* Analytical Chemistry Department, "Iuliu Haţieganu" University of Medicine and Pharmacy, 4 Pasteur Street, 400349, Cluj-Napoca, Romania
Abstract A hybrid graphene-polypyrrole (PPy) composite-based platform was elaborated by using nanosphere lithography structuration. This platform was further used for the antibody anti-acetaminophen immobilization in order to obtain an immunosensor for selective detection of paracetamol (acetaminophen) by using electrochemical methods. After patterning of the composite layer, the five-fold improvement of electrochemical signal was observed, suggesting that a higher amount of antibody was immobilized. This strategy permitted the ease and controlled immobilization of bioreceptors on the composite graphene-PPy platform and enhanced the sensitivity for paracetamol detection. © 2017 2016The TheAuthors. Authors.Published Published Elsevier © byby Elsevier Ltd.Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Peer-review under responsibility of the organizing committee of Biosensors 2016. Peer-review under responsibility of the organizing committee of Biosensors 2016 Keywords: graphene; polypyrrole; immunosensor; acetaminophen; nanosphere lithography
The entrapment of functionalized graphene in polypyrrole (PPy) conductive layers improves their engineering features. The association of graphene-PPy composite with nanosphere lithography generated a hybrid polystyrenegraphene-PPy patterned platform, with a further enhancement of their characteristics: an improved mechanical stability and electron transfer rate, combined with an increase of the active surface area >1, 2@. The immobilization of activated graphene oxide sheets onto the electrode was performed through electrostatic bonds, while the anti-paracetamol antibody (ApAb) biocomponent molecules were covalently bounded after the substrate modification with N-hydroxysuccinimide activated carboxylic functional groups. This strategy permits the controlled immobilization of ApAb in order to elaborate an immunosensor for paracetamol detection.
* Corresponding author. Tel.: +40 264 593118; fax: +40 264 597257. E-mail address: [email protected]
2212-0173 © 2017 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Peer-review under responsibility of the organizing committee of Biosensors 2016 doi:10.1016/j.protcy.2017.04.047
109
Andreea Cernat et al. / Procedia Technology 27 (2017) 108 – 109
The behavior of the nanostructured hybrid platform was tested by imagistic (Figure 1 B), impedimetric, and electrochemical quartz crystal microbalance experiments proving the successful patterning of the composite material and the biomolecule’s immobilization.
A
C B Fig. 1. (A) The development of the nanostructured platform and the immunosensor fabrication; (B) Optical microscopy images performed on the PS-graphene-PPy composite and (C) Current intensity variation, following the incubation with 5 µM paracetamol solution, recorded in PBS at the unstructured graphene-PPy (black line) and the nanostructured graphene-PPy (red line); (ApAb was incubated for 60 minutes; 5 % BSA for 20 minutes and paracetamol for 15 minutes) [3]
The unstructured and nanostructured graphene-PPy hybrid platforms were elaborated and the properties were tested by using paracetamol, a stable molecule with electrochemical activity. All the steps involved in immunosensor elaboration, presented in Figure 1 A, were optimized and it was observed that the sensor allowed the specific detection of paracetamol in a linear range of 0.1 to 25 µM at the unstructured platform. The analytical signal was of about 5 times improved on the patterned material in comparison with the unstructured one, proving the superior features of the structured surface (Figure 1 C). This nanopatterning strategy and the possibility of ease and controlled immobilization of bioreceptors may be exploited in biosensors development and applied in biomedical domain for the detection of biomarkers. Acknowledgements The authors are grateful for the financial support from the Romanian National Authority for Scientific Research, for project number PN-II-ID-PCE-2011-3-0355.
References [1] Cernat A, Griveau S, Martin P, Lacroix JC, Farcău C, Săndulescu R, Bedioui F. Electrografted nanostructured platforms for click chemistry. Electrochem Commun 2012;23:141-4. [2] Cernat A, Le Goff A, Holzinger M, Săndulescu R, Cosnier S. Micro-to nanostructured poly(pyrrole-nitrilotriacetic acid) films via nanosphere templates: applications to 3D enzyme attachment by affinity interactions. Anal Bioanal Chem 2014;406:1141-7. [3] Cernat A, Tertiş M, Păpară CN, Bodoki E, Săndulescu R. Nanostructured platform based on graphene-polypyrrole composite for immunosensor fabrication. Int J Electrochem Sci 2015;10:4718-31.