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Ultrasensitive electrochemical sensor for detection of salivary cortisol in stress conditions

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Abstract

A natural stress response induces elevated cortisol levels in biological fluids, such as saliva. While current sensor technologies can detect cortisol in real time, their sensitivity and reliability for human subjects have not been assured. This is due to relatively low concentrations of salivary cortisol, which fluctuate throughout the day and vary significantly between individuals. To address these challenges, we present an improved electrochemical biosensor leveraging graphene’s exceptional conductivity and physicochemical properties. A 1-pyrenebutyric acid N-hydroxysuccinimide ester (PBASE-NHS)–modified commercial graphene foam (GF) electrode is presented to realize an ultra-sensitive biosensor for cortisol detection directly in human saliva. The biosensor fabrication process entails the attachment of anti-cortisol monoclonal antibodies (mAb-cort) onto a PBASE-NHS/GF electrode through noncovalent immobilization on the vertically stratified graphene foam electrode surface. This unique immobilization strategy preserves graphene’s structural integrity and electrical conductivity while facilitating antibody immobilization. The binding of cortisol to immobilized mAb-cort is read out via differential pulse voltammetry using ferri/ferro redox reactions. The immunosensor demonstrates an exceptional dynamic range of 1.0 fg mL−1 to 10,000 pg mL−1 (R2 = 0.9914) with a detection limit of 0.24 fg mL−1 (n = 3) for cortisol. Furthermore, we have established the reliability of cortisol sensors in monitoring human saliva. We have also performed multiple modes of validation, one against the established enzyme-linked immunosorbent assay (ELISA) and a second by a third-party service Salimetric on 16 student volunteers exposed to different stress levels, showing excellent correlation (r = 0.9961). These findings suggest the potential for using mAb-cort/PBASE-NHS/GF-based cortisol electrodes for monitoring salivary cortisol in the general population.

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Funding

This work was supported by the National Science Foundation (NSF) under Grant 1931978. It was also partially supported through the TTW program at the Uniformed Services University of Health Sciences (USUHS) under the award HU0001-20-2-0014 and the DoD Peer Review Medical Research Program award number W81XWH-21-2-0012. Human samples were collected as per protocol approved by the Institutional Review Board (IRB) at Tufts University (1706030).

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Authors and Affiliations

  1. Nano Lab, Advanced Technology Laboratory, Tufts University, Medford, MA, 02155, USA

    Atul Sharma & Sameer Sonkusale

  2. Department of Electrical and Computer Engineering, Tufts University, Medford, MA, 02155, USA

    Atul Sharma & Sameer Sonkusale

  3. Department of Psychology, Tufts University, Medford, MA, 02155, USA

    Alia Wulff & Ayanna Thomas

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Sharma, A., Wulff, A., Thomas, A. et al. Ultrasensitive electrochemical sensor for detection of salivary cortisol in stress conditions. Microchim Acta 191, 103 (2024). https://doi.org/10.1007/s00604-023-06169-0

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