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Open AccessArticle

Electrochemical Immunosensor Based on Nanoelectrode Ensembles for the Serological Analysis of IgG-type Tissue Transglutaminase

1
Department of Molecular Sciences and Nanosystems, University Ca’Foscari of Venice, via Torino 155, 30172 Venezia Mestre, Italy
2
Institute for Maternal and Child Health - IRCCS “Burlo Garofolo”, 34100 Trieste, Italy
3
Department of Medical, Surgical and Health Sciences, University of Trieste, 34100 Trieste, Italy
*
Author to whom correspondence should be addressed.
Sensors 2019, 19(5), 1233; https://doi.org/10.3390/s19051233
Received: 22 December 2018 / Revised: 4 March 2019 / Accepted: 5 March 2019 / Published: 11 March 2019
(This article belongs to the Special Issue Nanostructured Surfaces in Sensing Systems)
Celiac disease (CD) is a gluten-dependent autoimmune disorder affecting a significant percentage of the general population, with increasing incidence particularly for children. Reliable analytical methods suitable for the serological diagnosis of the disorder are urgently required for performing both the early diagnosis and the follow-up of a patient adhering to a gluten-free diet. Herein we report on the preparation and application of a novel electrochemical immunosensor based on the use of ensembles of gold nanoelectrodes (NEEs) for the detection of anti-tissue transglutaminase (anti-tTG), which is considered one reliable serological marker for CD. To this end, we take advantage of the composite nature of the nanostructured surface of membrane-templated NEEs by functionalizing the polycarbonate surface of the track-etched membrane with tissue transglutaminase. Incubation of the functionalized NEE in anti-tTG samples results in the capture of the anti-tTG antibody. Confirmation of the recognition event is achieved by incubating the NEE with a secondary antibody labelled with horseradish peroxidase (HRP): in the presence of H2O2 as substrate and hydroquinone as redox mediator, an electrocatalytic current is indeed generated whose increment is proportional to the amount of anti-tTG captured from the sample. The optimized sensor allows a detection limit of 1.8 ng mL−1, with satisfactory selectivity and reproducibility. Analysis of serum samples from 28 individuals, some healthy and some affected by CD, furnished analytical results comparable with those achieved by classical fluoroenzyme immunoassay (FEIA). We note that the NEE-based immunosensor developed here detects the IgG isotype of anti-tTG, while FEIA detects the IgA isotype, which is not a suitable diagnostic marker for IgA-deficient patients. View Full-Text
Keywords: celiac disease; immunosensor; anti-tissue transglutaminase; nanoelectrode array; voltammetry celiac disease; immunosensor; anti-tissue transglutaminase; nanoelectrode array; voltammetry
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Habtamu, H.B.; Not, T.; De Leo, L.; Longo, S.; Moretto, L.M.; Ugo, P. Electrochemical Immunosensor Based on Nanoelectrode Ensembles for the Serological Analysis of IgG-type Tissue Transglutaminase. Sensors 2019, 19, 1233.

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