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Chitosan-coated polystyrene microplate for covalent immobilization of enzyme

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Abstract

Microplates made of polystyrene have been widely used for immunoassays. Protein molecules that have been immobilized on a hydrophobic polystyrene microplate by passive adsorption lose their activity and suffer considerable denaturation. A new chitosan-coated microplate suitable for the covalent immobilization of enzymes has been developed. The primary amino groups of chitosan were exploited for this covalent coupling of proteins. The optical transmittance of the chitosan-coated microplate, at wavelengths of 400–800 nm, was estimated to be suitable for its application in chromogenic reaction-based bioassays. The immobilization efficiency of the chitosan-coated microplate was demonstrated to be far superior to that of a conventional microplate when tested using acetylcholinesterase (AChE) and β-glucosidase as model biomolecules, and the chitosan-coated microplate may thus have potential applications in biosensing and bioreactor systems.

Comparison of the optical transmittances of chitosan-coated microplates in the wavelength range monitored in chromogenic reactions employed for immunoassays with the transmittance of a conventional polystyrene microplate

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (no. 30600494) and the Fundamental Research Fund for the Central Universities (no. GK200902010)

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Authors and Affiliations

  1. Key Laboratory of Applied Surface and Colloid Chemistry (Shaanxi Normal University), Ministry of Education, Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Materials Science, Shaanxi Normal University, Xi’an, 710062, China

    Yaodong Zhang, Li Li, Caihong Yu & Tingting Hei

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  1. Yaodong Zhang
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  2. Li Li
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  3. Caihong Yu
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  4. Tingting Hei
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Correspondence to Yaodong Zhang.

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Zhang, Y., Li, L., Yu, C. et al. Chitosan-coated polystyrene microplate for covalent immobilization of enzyme. Anal Bioanal Chem 401, 2311–2317 (2011). https://doi.org/10.1007/s00216-011-5306-7

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  • DOI: https://doi.org/10.1007/s00216-011-5306-7

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