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A novel reverse fluorescent immunoassay approach for sensing human chorionic gonadotropin based on silver-gold nano-alloy and magnetic nanoparticles

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Abstract

A novel and environmentally friendly reverse fluorescent immunoassay approach was proposed and utilized for sensing human chorionic gonadotropin (HCG) in human serum by coupling a newly prepared and highly fluorescent glutathione-stabilized silver-gold nano-alloy (GSH-AgAuNAs) with magnetic nanoparticles (MNPs). To construct such a reverse system, fluorescent GSH-AgAuNAs and MNPs were first prepared and bio-functionalized with monoclonal antibodies (Mab-I and Mab-II) toward HCG antigen, respectively. Then, the GSH-AgAuNAs functionalized with Mab-I were incubated with HCG, followed by the addition of MNPs attached to Mab-II. Thereafter, a sandwich-type immunoassay could be constructed for determination of HCG owing to the antibody-antigen recognition between the functionalized GSH-AgAuNAs and MNPs. Afterwards, a magnetic collection was employed. Hence, the amount of GSH-AgAuNAs would be reduced through an immuno-magnetic separation, thus weakening the fluorescent intensity. Different from conventional immunoassay, our work determined the quantitative signal by measuring the decreasing gradient fluorescent intensity. Under optimal conditions, the developed reverse method exhibited a wide linear range of 0.5–600 ng mL−1 toward HCG with a detection limit of 0.25 ng mL−1. Additionally, the proposed immunoassay was validated using spiked samples, illustrating a satisfactory result in practical application.

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Acknowledgments

We are grateful for the financial support from the Natural Science Foundation of Guangdong Province (No. S2011010005208 and No. 2014A030313480), the Science & Technology Project of Guangdong Province (No. 2013B030600001), and the Guangdong High Education Fund of Science and Technology Innovation (No. 2013KJCX0078).

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    Authors and Affiliations

    1. Department of Chemistry and Laboratory for Preparation and Application of Ordered Structural Materials of Guangdong Province, Shantou University, Shantou, Guangdong, 515063, China

      Xiaopeng Huang, Xiang Huang, Xiaona Xie & Wenhua Gao

    2. Department of Pharmacy, Taishan Medicine College, Taian, Shandong, 271016, China

      Yuqin Li

    3. Analysis & Testing Center, Shantou University, Shantou, Guangdong, 515063, China

      Yanping Xu, Yaowen Chen & Wenhua Gao

    Authors
    1. Xiaopeng Huang
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    2. Yuqin Li
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    3. Xiang Huang
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    5. Yanping Xu
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    6. Yaowen Chen
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    7. Wenhua Gao
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    Correspondence to Wenhua Gao.

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    3. A single study is not split up into several parts to increase the quantity of submissions and submitted to various journals or to one journal over time.

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    Xiaopeng Huang and Yuqin Li contributed equally to this work.

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    Huang, X., Li, Y., Huang, X. et al. A novel reverse fluorescent immunoassay approach for sensing human chorionic gonadotropin based on silver-gold nano-alloy and magnetic nanoparticles. Anal Bioanal Chem 408, 619–627 (2016). https://doi.org/10.1007/s00216-015-9144-x

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