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Screening for cystic fibrosis via a magnetic and microfluidic immunoassay format with electrochemical detection using a copper nanoparticle-modified gold electrode

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Abstract

This article describes a microfluidic electrochemical immunoassay that features two strategies, viz. (a), the incorporation of magnetic nanoparticles (MNPs) into the central microfluidic channel and acting as a bioaffinity support for the immobilization of the antibody against the immunoreactive trypsin (anti-IRT), and (b), the electrodeposition of copper nanoparticles (CuNPs) on a gold electrode. IRT, a marker for cystic fibrosis, is extracted from blood samples onto a disk using ultrasonication, eluted, and then injected into the detection system where it is captured by anti-IRT-loaded nanoparticles (anti-IRT-Ab-MNPs). Bound IRT is electrochemically quantified after addition of HRP-labeled anti-IRT-Ab which, in the presence of H2O2, catalyzes the oxidation of catechol to form o-benzoquinone which is detected at a working potential of −150 mV (vs. Ag/AgCl). The electrochemical response to benzoquinone is proportional to the concentration of IRT in the range from 0 to 580 ng⋅mL−1. The coefficients of variation are <5 % for within-day assays, and <6.4 % for between-day assays. The method was compared to a commercial ELISA for IRT where is showed a correlation coefficient of close to 1. In our perception, this approach represents an attractive alternative to existing methods for screening newborns for cystic fibrosis.

Immunoreactive trypsin (IRT) extracted from blood samples was captured into the central microfluidic channel by anti-IRT-loaded magnetic nanoparticles. Bound IRT was electrochemically quantified after addition of HRP-labeled anti-IRT-Ab using a gold electrode modified with copper nanoparticles.

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Acknowledgments

The authors wish to thank the financial support from the Universidad Nacional de San Luis (UNSL), the Instituto de Química de San Luis (INQUISAL), the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) and the Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT).

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  1. INQUISAL, Department of Chemistry, National University of San Luis, CONICET, Chacabuco 917. D5700BWS, San Luis, Argentina

    Maria Luz Scala Benuzzi, Sirley V. Pereira, Julio Raba & Germán A. Messina

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  1. Maria Luz Scala Benuzzi
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  2. Sirley V. Pereira
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  3. Julio Raba
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Correspondence to Germán A. Messina.

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Benuzzi, M.L.S., Pereira, S.V., Raba, J. et al. Screening for cystic fibrosis via a magnetic and microfluidic immunoassay format with electrochemical detection using a copper nanoparticle-modified gold electrode. Microchim Acta 183, 397–405 (2016). https://doi.org/10.1007/s00604-015-1660-z

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  • DOI: https://doi.org/10.1007/s00604-015-1660-z

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