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Comparison of monomeric and polymeric horseradish peroxidase as labels in competitive ELISA for small molecule detection

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Abstract

We have developed a simple and sensitive competitive enzyme-linked immunosorbent assay (ELISA) to determine aflatoxin B1 (as a model small analyte) and using streptavidin-polymeric horseradish peroxidase complex (SApolyHRP) as a label for signal amplification. The performance of the assay was evaluated by comparing it with the classical indirect competitive ELISA using HRP labeled anti-mouse IgG as the tracer antibody. The results indicate that the SApolyHRP-based competitive ELISA exhibits a typically 2.4-fold steeper slope of the linear working range of the calibration curve compared to the monomeric HRP based classical ELISA, i.e., the sensitivity was increased. The SApolyHRP conjugate causes a typically 19-fold stronger signal generation in comparison to the traditional HRP labeled anti-mouse IgG at the same concentration (25 ng mL−1). Moreover, the SApolyHRP-based assay has a much wider linear range and a 3.8-fold better signal-to-noise ratio. Considering its simplicity, sensitivity and ease of operation, this competitive ELISA is considered to be a promising tool for small molecule immunodetection.

We have developed a polymeric horseradish peroxidase (polyHRP) based competitive ELISA for the detection of small molecule (aflatoxin B1 as model) and have compared ELISAs using polyHRP and monomeric HRP as labels in terms of analytical performance, i.e., sensitivity, signal-to-noise, linear range and analysis time.

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Acknowledgements

This research was supported by the National Natural Science Foundation of China (No. 30825027) and German Ministry of Economics and Technology (via AiF) and the FEI (Forschungskreis der Ernährungsindustrie e.V. Bonn); project AiF 381 ZN. D. Li would like to thank National Public Welfare Project for Agriculture (No. 200903009) and Graduate Student International Exchange Fund of Zhejiang University for supporting his stay at the Technische Universität München.

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Authors and Affiliations

  1. College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, People’s Republic of China

    Dongyang Li, Yibin Ying & Jian Wu

  2. Institute of Hydrochemistry & Chair of Analytical Chemistry, Technische Universität München, Marchioninistrasse 17, 81377, Munich, Germany

    Dongyang Li, Reinhard Niessner & Dietmar Knopp

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  1. Dongyang Li
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  2. Yibin Ying
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  3. Jian Wu
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  4. Reinhard Niessner
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  5. Dietmar Knopp
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Correspondence to Yibin Ying or Dietmar Knopp.

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Li, D., Ying, Y., Wu, J. et al. Comparison of monomeric and polymeric horseradish peroxidase as labels in competitive ELISA for small molecule detection. Microchim Acta 180, 711–717 (2013). https://doi.org/10.1007/s00604-013-0974-y

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  • DOI: https://doi.org/10.1007/s00604-013-0974-y

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