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Electrochemical immunoassay for one-pot detection of thyroxin (T4) and thyroid-stimulating hormone (TSH) using magnetic and Janus nanoparticles

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Abstract

Concurrent measurement of thyroid-stimulating hormone (TSH) and thyroxine (T4) hormones profoundly help clinicians diagnose hyper- and hypothyroidism. This work demonstrates the development of a sandwich-type electrochemical immunoassay using Janus and magnetic nanoparticles for one-pot detection of thyroxine (T4) and thyroid-stimulating hormone (TSH). The signaling probe was developed by preparing Janus cadmium (CdO) and zinc oxide (ZnO) NPs decorated by T4/TSH-specific molecularly imprinted polymers (MIPT4-CdO and MIPTSH-ZnO). The capture probe was obtained by coating magnetic Fe3O4 NPs with 1,3-Bis(3-carboxy propyl) tetramethyl disiloxane and activating using N-hydroxy succinimide (NHS) and 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC), and finally conjugating with T4/TSH-specific antibodies. To analyze T4 and TSH in actual samples, MIPT4-CdO and MIPTSH-ZnO were added to the sample solutions, and after incubation, capture probes (Fe3O4-AbTSH and Fe3O4-AbT4) were added. An external magnetic field was used to separate the sandwiched nanosystem, followed by adding a dilute solution of nitric acid (HNO3) to dissolve CdO and ZnO NPs and free Cd(II) and Zn(II) cations. The concentration of these cations was determined using constant-current potentiometric stripping analysis (cc-PSA) on screen-printed electrodes (SPE) modified with multi-walled carbon nanotubes (MWCNT). The obtained signals for Cd(II) and Zn(II) were proportional to T4 and TSH concentrations. Limits of detection (LOD) for T4 and TSH analyses were respectively 0.02 ng.dL−1 and 0.0002 µU.mL−1 with a linear range of 0.05–50 ng.dL−1 and 0.001–100 µU.mL−1. The proposed nanosystem’s main advantage is the simultaneous detection of T4 and TSH in clinical samples with high sensitivity, selectivity, and stability.

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Funding

The Biosensor Sciences and Technologies Research Center, Ardabil University of Medical Sciences, Ardabil, Iran, financially supported the current study (Grant No.:1002756).

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Authors and Affiliations

  1. Biosensor Sciences and Technologies Research Center, Ardabil University of Medical Sciences, Ardabil, Iran

    Pari Karami & Mohammad Johari-Ahar

  2. Department of Medicinal Chemistry, School of Pharmacy, Ardabil University of Medical Sciences, Ardabil, Iran

    Danial Gholamin & Mohammad Johari-Ahar

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  1. Pari Karami
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  2. Danial Gholamin
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Contributions

Mohammad Johari-Ahar: conceptualization, methodology, funding acquisition, writing—review and editing, supervision.

Pari Karami: methodology, investigation, formal analysis, writing—review, and editing.

Danial Gholamin: investigation and formal analysis.

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Correspondence to Mohammad Johari-Ahar.

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The protocols for this study have been approved by the ethics committee of Ardabil University of Medical Sciences, Ardabil, Iran (Approval No: IR.ARUMS.REC.1398.614).

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Karami, P., Gholamin, D. & Johari-Ahar, M. Electrochemical immunoassay for one-pot detection of thyroxin (T4) and thyroid-stimulating hormone (TSH) using magnetic and Janus nanoparticles. Anal Bioanal Chem 415, 4741–4751 (2023). https://doi.org/10.1007/s00216-023-04767-8

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