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Electrochemical sandwich immunoassay for the prostate specific antigen using a polyclonal antibody conjugated to thionine and horseradish peroxidase

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Abstract

The authors present a voltammetric immunoassay for the quantification of the prostate specific antigen (PSA), a well-established tumor marker. Biotinylated anti- PSA antibodies were immobilized on the surface of a nano-TiO2-modified carbon paste electrode using specific interaction between streptavidin and biotin. Streptavidin itself was covalently grafted to the electrode surface via 4-aminobenzoic acid film. Signal transduction was then performed using polyclonal antibodies conjugated with horseradish peroxidase (HRP) and thionine. An amplified catalytic reduction current was observed in the presence of H2O2 because more than one polyclonal antibody linked to each antigen. Each step of the preparation of the immunosensor was monitored via electrochemical impedance spectroscopy. The electrode, if operated at a typical potential of 0.3 V vs. Ag/AgCl and using hexacyanoferrate as an electrochemical probe, exhibits linear responses in the 0.10 to 5.0 ng·mL−1 and 5.0 to 100 ng·mL−1 PSA concentration ranges, with a detection limit of 200 pg·mL−1. The accuracy of the biosensor was confirmed by analyzing a certified human serum sample, and this indicated that the immunosensor is well suited for the quantification of PSA.

Schematic presentation of a sandwich-type electrochemical immunosensor for the prostate specific antigen. It is based on the use of a carbon paste electrode, streptavidin and biotinylated monoclonal antibody. Signal transduction is the result of the catalytic reduction of thionine.

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Authors and Affiliations

  1. Department of Chemistry, Shahid Bahonar University of Kerman, Kerman, Iran

    Zahra Biniaz, Ali Mostafavi & Tayebeh Shamspur

  2. Department of Biotechnology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran

    Masoud Torkzadeh-Mahani

  3. Pistachio Safety Research Center, Rafsanjan University of Medical Sciences, Rafsanjan, Iran

    Maryam Mohamadi

Authors
  1. Zahra Biniaz
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  2. Ali Mostafavi
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  3. Tayebeh Shamspur
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  4. Masoud Torkzadeh-Mahani
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  5. Maryam Mohamadi
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Correspondence to Tayebeh Shamspur or Maryam Mohamadi.

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Biniaz, Z., Mostafavi, A., Shamspur, T. et al. Electrochemical sandwich immunoassay for the prostate specific antigen using a polyclonal antibody conjugated to thionine and horseradish peroxidase. Microchim Acta 184, 2731–2738 (2017). https://doi.org/10.1007/s00604-017-2284-2

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  • DOI: https://doi.org/10.1007/s00604-017-2284-2

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