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Gold nanoparticle-labeled detection antibodies for use in an enhanced electrochemical immunoassay of hepatitis B surface antigen in human serum

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Abstract

The application of gold nanoparticle-based electrochemical immunoassays have been extensively studied for the detection of hepatitis B surface antigen (HBsAg), but most often they exhibit low sensitivity. We describe the fabrication of a new electrochemical immunoassay for signal amplification of the antigen-antibody reaction combined with the nanogold-based bio-barcode technique. Hepatitis B surface antibody (HBsAb) was initially immobilized on a nanogold/thionine/DNA-modified gold electrode, and then a sandwich-type immunoassay format was employed for the detection of HBsAg using nanogold-codified horseradish peroxidase-HBsAb conjugates as secondary antibodies. Under optimal conditions, the current response of the sandwich-type immunocomplex relative to the H2O2 system was proportional to HBsAg concentration in the range from 0.5 to 650 ng·mL−1 with a detection limit of 0.1 ng·mL−1 (S/N = 3). The precision, reproducibility and stability of the immunosensor were acceptable. Subsequently, the immunosensors were used to assay HBsAg in human serum specimens. Analytical results were in agreement with those obtained by the standard chemiluminescence enzyme-linked immunosorbent assay.

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Acknowledgements

This project was funded by the National Natural Science Foundation of China (No.30670628) and Military Medical Research Foundation of China (06MA188).

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Authors and Affiliations

  1. Department of Hepatobiliary Surgery, 101th Hospital of Chinese People’s Liberation Army, Wuxi, 214044, PR China

    Sheng Wu

  2. Department of Cancer Center, Research Institute of Surgery, Daping Hospital, Third Military Medical University, Chongqing, 400042, PR China

    Zhaoyang Zhong, Dong Wang, Mengxia Li, Yi Qing, Nan Dai & Zengpeng Li

Authors
  1. Sheng Wu
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  2. Zhaoyang Zhong
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  3. Dong Wang
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  4. Mengxia Li
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  5. Yi Qing
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  6. Nan Dai
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  7. Zengpeng Li
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Corresponding authors

Correspondence to Sheng Wu or Dong Wang.

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Wu and Zhong contributed equally to this work.

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Wu, S., Zhong, Z., Wang, D. et al. Gold nanoparticle-labeled detection antibodies for use in an enhanced electrochemical immunoassay of hepatitis B surface antigen in human serum. Microchim Acta 166, 269–275 (2009). https://doi.org/10.1007/s00604-009-0184-9

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  • DOI: https://doi.org/10.1007/s00604-009-0184-9

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