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An immunoassay using antibody-gold nanoparticle conjugate, silver enhancement and flatbed scanner

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Abstract

This study reports a convenient immunoassay using antibody-gold nanoparticle (Ab-AuNP) conjugate as a reporter molecule and a flatbed scanner for the optical scanning and measuring of the immuno-reaction signal. The silver enhancement reaction, a signal amplification method in which silver ions are reduced to silver metal, is introduced to magnify the detection signal. The whole framework of the study is based on (1) the direct immunoassay (two-layer format) in which the antigen is directly immobilized on the chip, and (2) the sandwich immunoassay [three-layer format, process similar to enzyme-linked immunosorbent assay (ELISA)], which contains a primary antibody, a secondary antibody (antibody-nanoparticle conjugate) and antigens. The experimental data show that the micro-scale silver precipitation phenomenon is catalyzed by Ab-AuNP conjugates. This reaction can be observed by the naked eye, even at an IgG-AuNPs concentration of 5 pM. The relationships between sample concentration and detection signal are discussed, and the detection limit (sandwich assay) for the sample antigen is 1 ng/mL. Using a flatbed scanner, Ab-AuNP conjugates and a silver enhancement reaction, a new immunoassay is constructed.

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Acknowledgments

The authors would like to thank the Center for Micro/Nano Technology, National Cheng Kung University, Tainan for access to their equipment and for their technical support. Funding from the Ministry of Education and the National Science Council of Taiwan, ROC under contract number (NSC 95-2323-B-006-005, NSC 95-2323-B-006-006) is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Engineering Science, National Cheng Kung University, 1 University Road, Tainan, 701, Taiwan, ROC

    Chia-Hsien Yeh, Ching-Yuan Hung & Yu-Cheng Lin

  2. Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, Taiwan, ROC

    Tsung Chain Chang

  3. Department of Chemistry, National Cheng Kung University, Tainan, Taiwan, ROC

    Hong-Ping Lin

  4. Center for Micro/Nano Technology, National Cheng Kung University, Tainan, Taiwan, ROC

    Yu-Cheng Lin

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  1. Chia-Hsien Yeh
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  2. Ching-Yuan Hung
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  3. Tsung Chain Chang
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  4. Hong-Ping Lin
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  5. Yu-Cheng Lin
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Correspondence to Yu-Cheng Lin.

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Yeh, CH., Hung, CY., Chang, T.C. et al. An immunoassay using antibody-gold nanoparticle conjugate, silver enhancement and flatbed scanner. Microfluid Nanofluid 6, 85–91 (2009). https://doi.org/10.1007/s10404-008-0298-0

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  • DOI: https://doi.org/10.1007/s10404-008-0298-0

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