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Label-free C-reactive protein electronic detection with an electrolyte-gated organic field-effect transistor-based immunosensor

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Abstract

In this contribution, we propose a label-free immunosensor, based on a novel type of electrolyte-gated field-effect transistor (EGOFET), for ultrasensitive detection of the C-reactive protein (CRP). The recognition layer of the biosensor is fabricated by physical adsorption of the anti-CRP monoclonal antibody onto a poly-3-hexyl thiophene (P3HT) organic semiconductor surface. A supplementary nonionic hydrophilic polymer is used as a blocking agent preventing nonspecific interactions and allowing a better orientation of the antibodies immobilized onto the P3HT surface. The whole biomolecule immobilization procedure does not require any pretreatment of the organic semiconductor surface, and the whole functionalization process is completed in less than 30 min. Surface plasmon resonance (SPR) measurements were performed to assess the amount of biomolecules physisorbed onto the P3HT and to evaluate the CRP binding proprieties of the deposited anti-CRP layer. A partial surface coverage of about 23 % of adsorbed antibody molecules was found to most efficiently sense the CRP. The electrical performance of the EGOFET immunosensor was comparable to that of a bare P3HT EGOFET device, and the obtained CRP calibration curve was linear over six orders of magnitude (from 4 pM to 2 μM). The relative standard deviation of the individual calibration points, measured on immunosensors fabricated on different chips, ranged between 1 and 14 %, and a detection limit of 2 pM (220 ng/L) was established. The novel electronic immunosensor is compatible with low-cost fabrication procedures and was successfully employed for the detection of the CRP biomarker in the clinically relevant matrix serum.

Schematic of the EGOFET immunosensor for CRP detection. The anti-CRP monoclonal antibody layer is physisorbed on the P3HT organic semiconductor and the CRP is directly measured by a label-free electronic EGOFET transducer

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Acknowledgments

The “Electrolyte-Gated Organic Field-Effect Biosensors—BioEGOFET” project FP7/2007-2013 under Grant agreement no. 248728, the “OFET biosensors for point-of-care applications–sense-of-care” project, FP7-People-ITN-EID-2012 under Grant Agreement 316845 and the PON project “Laboratorio per lo Sviluppo Integrato delle Scienze e delle Tecnologie dei Materiali Avanzati e per dispositivi innovativi-LABORATORIO SISTEMA,” and the PRIN 2010–2011 project no. 2010BJ23MN “Nanostructured Soft Matter” by the Italian MIUR (Ministry of Education, Universities and Research) are acknowledged for partial financial support of this work. Professor Gaetano Scamarcio is acknowledged for useful discussions. Dr. Cinzia Di Franco is acknowledged for her support in the realization of photo-lithographically patterned substrates.

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Authors and Affiliations

  1. Dipartimento di Chimica, Università degli Studi di Bari “Aldo Moro” and GSI, Via Orabona 4, 70126, Bari, Italy

    Maria Magliulo, Donato De Tullio, Kyriaki Manoli, Gerardo Palazzo & Luisa Torsi

  2. BioMediTech, University of Tampere, Fimlab Laboratories Ltd., 33520, Tampere, Finland

    Inger Vikholm-Lundin

  3. BioNavis OY, Elopellontie 3.C, 33470, Ylöjärvi, Finland

    Willem M. Albers

  4. VTT Technical Research Centre of Finland, Process Chemistry and Environmental Engineering, Sinitaival 6, 33720, Tampere, Finland

    Tony Munter

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  1. Maria Magliulo
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  2. Donato De Tullio
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  3. Inger Vikholm-Lundin
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  4. Willem M. Albers
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  6. Kyriaki Manoli
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Corresponding author

Correspondence to Maria Magliulo.

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The study has been approved by the ethics committee of the University of Bari “Aldo Moro” and has been performed in accordance with the ethical standards.

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The authors declare that they have no conflict of interest.

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Published in the topical collection featuring Young Investigators in Analytical and Bioanalytical Science with guest editors S. Daunert, A. Baeumner, S. Deo, J. Ruiz Encinar, and L. Zhang.

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Magliulo, M., De Tullio, D., Vikholm-Lundin, I. et al. Label-free C-reactive protein electronic detection with an electrolyte-gated organic field-effect transistor-based immunosensor. Anal Bioanal Chem 408, 3943–3952 (2016). https://doi.org/10.1007/s00216-016-9502-3

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