期刊
TALANTA
卷 211, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.talanta.2020.120761
关键词
Interferon gamma; IFN-gamma; Electrochemical immunosensor; Aryl diazonium salt; Grafting; Saliva
资金
- Spanish Ministerio de Economia y Competitividad [CTQ2015-64402-C2-1-R]
- Spanish Ministerio de Ciencia, Innovacion y Universidades [RTI2018-096135-B-I00]
- TRANSNANOAVANSENS-CM Program from the Comunidad de Madrid [S2018/NMT-4349]
A simple, fast and sensitive amperometric immunosensing method for the determination of the clinically relevant cytokine interferon gamma (IFN-gamma) in saliva complying the requirements demanded for this kind of sample is reported. The target analyte was sandwiched between a specific capture antibody covalently immobilized on a screen-printed electrode functionalized by the diazonium salt grafting of p-aminobenzoic acid, and a biotinylated detector antibody labeled with a streptavidin-horseradish peroxidase conjugate. The amperometric responses measured at - 0.20 V vs Ag pseudo-reference electrode upon addition of hydrogen peroxide in the presence of hydroquinone as the redox mediator allowed a calibration plot with a linear range between 2.5 and 2000 pg mL(-1) and a low limit of detection (1.6 pg mL(-1)) to be obtained. In addition, a good selectivity against other non-target proteins was achieved. The developed method was validated by analyzing a WHO 1st International Standard for IFN-gamma. In addition, the immunosensor was used for the determination of the endogenous IFN-gamma in saliva with results in excellent agreement with those obtained by a commercial ELISA kit.
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