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Reagentless biosensor based on layer-by-layer assembly of functional multiwall carbon nanotubes and enzyme-mediator biocomposite

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Abstract

A simple and controllable layer-by-layer (LBL) assembly method was proposed for the construction of reagentless biosensors based on electrostatic interaction between functional multiwall carbon nanotubes (MWNTs) and enzyme-mediator biocomposites. The carboxylated MWNTs were wrapped with polycations poly(allylamine hydrochloride) (PAH) and the resulting PAH-MWNTs were well dispersed and positively charged. As a water-soluble dye methylene blue (MB) could mix well with horseradish peroxidase (HRP) to form a biocompatible and negativelycharged HRP-MB biocomposite. A (PAH-MWNTs/HRP-MB) n bionanomultilayer was then prepared by electrostatic LBL assembly of PAH-MWNTs and HRP-MB on a polyelectrolyte precursor film-modified Au electrode. Due to the excellent biocompatibility of HRP-MB biocomposite and the uniform LBL assembly, the immobilized HRP could retain its natural bioactivity and MB could efficiently shuttle electrons between HRP and the electrode. The incorporation of MWNTs in the bionanomultilayer enhanced the surface coverage concentration of the electroactive enzyme and increased the catalytic current response of the electrode. The proposed biosensor displayed a fast response (2 s) to hydrogen peroxide with a low detection limit of 2.0×10−7 mol/L (S/N=3). This work provided a versatile platform in the further development of reagentless biosensors.

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Authors and Affiliations

  1. Department of Chemistry, Zhejiang University, Hangzhou, 310027, China

    Xing-hua Zhou, Feng-na Xi, Yi-ming Zhang & Xian-fu Lin

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  1. Xing-hua Zhou
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  2. Feng-na Xi
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  3. Yi-ming Zhang
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  4. Xian-fu Lin
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Correspondence to Xian-fu Lin.

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Project (Nos. 20805043 and 30800247) supported by the National Natural Science Foundation of China

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Zhou, Xh., Xi, Fn., Zhang, Ym. et al. Reagentless biosensor based on layer-by-layer assembly of functional multiwall carbon nanotubes and enzyme-mediator biocomposite. J. Zhejiang Univ. Sci. B 12, 468–476 (2011). https://doi.org/10.1631/jzus.B1000280

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  • DOI: https://doi.org/10.1631/jzus.B1000280

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