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A nanohybrid composed of Prussian Blue and graphitic C3N4 nanosheets as the signal-generating tag in an enzyme-free electrochemical immunoassay for the neutrophil gelatinase-associated lipocalin

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Abstract

An enzyme-free electrochemical immunoassay is described for the neutrophil gelatinase-associated lipocalin (NGAL; a biomarker of kidney disease). Prussian Blue (PB) nanoparticles with redox activity were deposited on graphitic C3N4 nanosheets (g-C3N4) by in-situ reduction. A screen printed electrode (SPCE) was modified with antibody against NGAL, and the PB-g-C3N4 nanohybrid was used as the signal-generating tag for the secondary antibody against NGAL. Upon addition of target NGAL and of secondary antibody, a sandwich is formed on the SPCE. At an applied potential of typically 0.13 V (vs. Ag/AgCl), a well-defined voltammetric peak is observed that results from the presence of PB on the secondary antibody. Under optimal conditions, the peak current increases linearly in the 0.01 to 10 ng·mL−1 NGAL concentration range, and the detection limit is 2.8 pg·mL−1. An average precision of <12% was accomplished in the batch-to-batch mode. Other disease-related biomarkers do not interfere. The accuracy and inter-laboratory validation of this method were evaluated for target NGAL detection in spiked human serum by using a commercial ELISA. The results obtained by the two methods are in good accordance.

An enzyme-free electrochemical immunoassay was used for detection of neutrophil gelatinase-associated lipocalin by Prussian blut/graphitic-C3N4 nanohybrids as the signal-generation tags.

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Funding

Support by the Health Science Research Personnel Training Program of Fujian Province (Grant no.: 2017-CXB-22) and the Scientific Research Innovation Team Construction Program of Fujian Normal University (Grant no.: IRTL1702) is gratefully acknowledged.

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Author notes

  1. Fengling Zhang and Hongbin Zhong contributed equally to this work.

Authors and Affiliations

  1. Xiang’an Branch, The First Affiliated Hospital of Xiamen University, Xiamen University, Xiamen, 361003, China

    Fengling Zhang, Hongbin Zhong, Ying Lin & Miaoxuan Chen

  2. Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, College of Life Sciences, Fujian Normal University, Qishan Campus, Fuzhou, 350117, China

    Qingshui Wang & Yao Lin

  3. Medical College of Xiamen University, Xiang’an Branch, The First Affiliated Hospital of Xiamen University, Xiamen University, Xiamen, 361003, China

    Jiyi Huang

Authors
  1. Fengling Zhang
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  2. Hongbin Zhong
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  3. Ying Lin
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  4. Miaoxuan Chen
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  5. Qingshui Wang
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  6. Yao Lin
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Correspondence to Yao Lin or Jiyi Huang.

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Zhang, F., Zhong, H., Lin, Y. et al. A nanohybrid composed of Prussian Blue and graphitic C3N4 nanosheets as the signal-generating tag in an enzyme-free electrochemical immunoassay for the neutrophil gelatinase-associated lipocalin. Microchim Acta 185, 327 (2018). https://doi.org/10.1007/s00604-018-2865-8

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  • DOI: https://doi.org/10.1007/s00604-018-2865-8

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