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Label-free amperometric immunobiosensor based on a gold colloid and Prussian blue nanocomposite film modified carbon ionic liquid electrode

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Abstract

A novel experimental methodology based on a Prussian blue (PB) and gold nanoparticles (AuNPs) modified carbon ionic liquid electrode (CILE) was developed for use in a label-free amperometric immunosensor for the sensitive detection of human immunoglobulin G (HIgG) as a model protein. The CILE was fabricated by using the ionic liquid 1-octyl-3-methylimidazolium hexafluorophosphate as binder. Controllable electrodeposition of PB on the surface of the CILE and coating with 3-aminopropyl triethylene silane (APS) formed a film with high electronic catalytic activity and large surface area for the assembly of AuNPs and further immobilization of HIgG antibody. The electrochemistry of the formed nanocomposite biofilm was investigated by electrochemical techniques including cyclic voltammetry, differential pulse voltammetry, and electrochemical impedance spectroscopy. The HIgG concentration was measured through the decrease of amperometric responses in the corresponding specific binding of antigen and antibody. The decreased differential pulse voltammetric values were proportional to the HIgG concentration in two ranges, 0.05–1.25 ng mL−1 and 1.25–40 ng mL−1, with a detection limit of 0.001 ng mL−1 (S/N = 3). This electrochemical immunoassay combined the specificity of the immunological reaction with the sensitivity of the AuNPs, ionic liquid, and PB amplified electrochemical detection and would therefore be valuable for clinical immunoassays.

Cyclic voltammograms of the immunosensor in pH 7.0 phosphate-buffered solution containing 0.1 M KCl at the different scan rate of (from inner to outer) 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, and 800 mVs−1. The inset shows the linear relationship between the peak currents and the square root of scan rate.

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Acknowledgments

This work was supported by National Natural Science Foundation of China (20805040, 20635020), Excellent Youth Foundation of He’nan Scientific Committee (104100510020), China Postdoctoral Science Foundation funded project (200902508, 20080430163), Jiangsu Planned Projects for Postdoctoral Research Funds (0801041B), and sponsored by Program for Science & Technology Innovation Talents in Universities of Henan Province (2010HASTIT025).

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Authors and Affiliations

  1. Key Laboratory of Analytical Chemistry for Life Science (Ministry of Education of China), School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210093, People’s Republic of China

    Ke-Jing Huang & Jun-Jie Zhu

  2. College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang, 464000, People’s Republic of China

    Ke-Jing Huang, De-Jun Niu, Jun-Yong Sun & Xiao-Li Zhu

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  1. Ke-Jing Huang
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Correspondence to Jun-Jie Zhu.

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Huang, KJ., Niu, DJ., Sun, JY. et al. Label-free amperometric immunobiosensor based on a gold colloid and Prussian blue nanocomposite film modified carbon ionic liquid electrode. Anal Bioanal Chem 397, 3553–3561 (2010). https://doi.org/10.1007/s00216-010-3868-4

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