Simultaneous amperometric immunosensing of the metastasis-related biomarkers IL-13Rα2 and CDH-17 by using grafted screen-printed electrodes and a composite prepared from quantum dots and carbon nanotubes for signal amplification
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This paper describes a dual electrochemical immunoassay for the simultaneous determination of IL-13Rα2 and CDH-17, two biomarkers of emerging relevance in metastatic processes. The sandwich assay uses a screen-printed dual carbon electrode that was electrochemically grafted with p-aminobenzoic acid to allow the covalent immobilization of capture antibodies. A hybrid composed of graphene quantum dots (GQDs) and multiwalled carbon nanotubes (MWCNTs) act as nanocarriers for the detection antibodies and horseradish peroxidase. The use of this hybrid material considerably improves the assay (in comparison to the use of MWCNTs) due to the peroxidase mimicking activity of the GQDs. The method works at a low working potential (0.20 V vs. Ag pseudo-reference electrode) and thus is not readily interfered by unknown electroactive species. The dual immunoassay allows for the selective determination of both biomarkers with LOD values of 1.4 (IL-13sRα2) and 0.03 ng mL−1 (CDH-17). The simultaneous determination of IL-13Rα2 and CDH-17 was accomplished in lysates from breast and colorectal cancer cells with different metastatic potential, and in paraffin-embedded tumor tissues extracts from patients diagnosed with colorectal cancer at different stages. The applicability to discriminate the metastatic potential even in intact cells through the detection of both extracellular receptors has been demonstrated also. The assay can be performed within 3 h, requires small sample amounts (0.5 μg), and has a simple protocol.
KeywordsDual immunosensor Metastasis-related biomarkers Graphene quantum dots Signal amplification Electrochemical immunosensor Colorectal cancer Paraffin-embedded tissues Peroxidase mimic
The financial support of the Spanish Ministerio de Economía y Competitividad, Grants RTI2018-096135-B-I00 and CTQ2015-64402-C2-1-R, and the TRANSNANOAVANSENS-CM Program from the Comunidad de Madrid (Grant P2018/NMT-4349), are gratefully acknowledged. R.B. acknowledges the financial support of the PI17CIII/00045 Grant from the AES-ISCIII program. M.G.-A. was supported by a contract of the Programa Operativo de Empleo Juvenil y la Iniciativa de Empleo Juvenil (YEI) with the participation of the Consejería de Educación, Juventud y Deporte de la Comunidad de Madrid y del Fondo Social Europeo.
Compliance with ethical standards
The author(s) declare that they have no competing interests. This study and all the experimental protocols used were performed according to the guidelines and regulations and approved by the University Complutense of Madrid. The Institutional Ethical Review Board of the Hospital La Paz (Madrid, Spain), that supplied us with the clinical samples, approved this study and all patients given their written informed consent.
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