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Enzyme biosensor based on the immobilization of HRP on SiO2/BSA/Au composite nanoparticles

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Abstract

In this work, an enzyme biosensor based on the immobilization of horseradish peroxidase (HRP) on SiO2/BSA/Au/thionine/nafion-modified gold electrode was fabricated successfully. Firstly, nafion was dropped on the surface of the gold electrode to form a nafion film followed by chemisorption of thionine (Thi) as an electron mediator via the ion-exchange interaction between the Thi and nafion. Subsequently, the SiO2/BSA/Au composite nanoparticles were assembled onto Thi film through the covalent bounding with the amino groups of Thi. Finally, HRP was immobilized on the SiO2/BSA/Au composite nanoparticles due to the covalent conjugation to construct an enzyme biosensor. The surface topographies of the SiO2/BSA/Au composite nanoparticles were investigated by using scanning electronic microscopy. The stepwise self-assemble procedure of the biosensor was further characterized by means of cyclic voltammetry and chronoamperometry. The enzyme biosensor showed high sensitivity, good stability and selectivity, a wide linear response to hydrogen peroxide (H2O2) in the range of 8.0 × 10-6 ∼ 3.72 × 10-3 mol/L, with a detection limit of 2.0 × 10-6 mol/L. The Michaelies-Menten constant \( K_M^{app} \) value was estimated to be 2.3 mM.

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Abbreviations

v :

Scan rate(mV s-1)

E :

Potential (V)

I :

Current (µA)

C :

Concentration(mM)

T :

Time (s)

References

  1. Wan, J., Bi, J. L., Du, P., & Zhang, S. S. (2009). Analytical Biochemistry, 386, 256–261.

    Article  CAS  Google Scholar 

  2. Lupu, A., Lisboa, P., Valsesia, A., Colpo, P., & Rossi, F. (2009). Sensors and Actuators B, 137, 56–61.

    Article  Google Scholar 

  3. Li, F., Chen, W., Tang, C. F., & Zhang, S. S. (2009). Talanta, 77, 1304–1308.

    Article  CAS  Google Scholar 

  4. Camacho, C., Chico, B., & Cao, R. (2009). Biosensors & Bioelectronics, 24, 2028–2033.

    Article  CAS  Google Scholar 

  5. Gopalan, A. I., Lee, K.-P., & Ragupathy, D. (2009). Biosensors & Bioelectronics, 24, 2211–2217.

    Article  CAS  Google Scholar 

  6. Shimoumura, T., Itoh, T., Sumiya, T., Mizukami, F., & Ono, M. (2009). Talanta, 78, 217–220.

    Article  Google Scholar 

  7. Pingarron, J. M., Yanez-Sedeno, P., & Gonzalez-Cortes, A. (2008). Electrochimica Acta, 53, 5848–5866.

    Article  CAS  Google Scholar 

  8. Pang, X. Y., He, D. M., Luo, S. L., & Cai, Q. Y. (2009). Sensors and Actuators B, 137, 134–138.

    Article  Google Scholar 

  9. Kang, X. H., Wang, J., & Tang, Z. W. (2009). Talanta, 78, 120–125.

    Article  CAS  Google Scholar 

  10. Guo, S. J., & Dong, S. J. (2009). Trend in Analytical Chemistry, 28, 96–109.

    Article  CAS  Google Scholar 

  11. Zhu, Y. H., Cao, H. M., Tang, L. H., Yang, X. L., & Li, C. Z. (2009). Electrochimica Acta, 54, 2823–2827.

    Article  CAS  Google Scholar 

  12. Zhuo, Y., Yu, R. J., Yuan, R., & Chai, Y. Q. (2009). Journal of Electroanalytical Chemistry, 628, 90–96.

    Article  CAS  Google Scholar 

  13. Libor, Z., & Zhang, Q. (2009). Materials Chemistry and Physics, 114, 902–907.

    Article  CAS  Google Scholar 

  14. Kerman, K., Saito, M., Yamamura, S., & Takamura, Y. (2008). Trends in Analytical Chemistry, 27, 585–592.

    Article  CAS  Google Scholar 

  15. Zhu, A. W., Tian, Y., Liu, H. Q., & Luo, Y. P. (2009). Biomaterials, 30, 3183–3188.

    Article  CAS  Google Scholar 

  16. Shimomura, T., Itoh, T., Sumiya, T., Mizukami, F., & Ono, M. (2008). Sensors and Actuators B, 135, 268–275.

    Article  Google Scholar 

  17. Di, J. W., Shen, C. P., & Peng, S. H. (2005). Analytica Chimica Acta, 553, 196–200.

    Article  CAS  Google Scholar 

  18. Zhang, Y. Z., Zhang, K. Y., & Ma, H. Y. (2009). Analytical Biochemistry, 387, 13–19.

    Article  CAS  Google Scholar 

  19. Wang, Y. Y., Chen, X. J., & Zhu, J. J. (2009). Electrochemistry Communications, 11, 323–326.

    Article  CAS  Google Scholar 

  20. Lamarre, J.-M., Billard, F., Kerboua, C. H., & Lequime, M. (2008). Optics Communications, 281, 331–340.

    Article  CAS  Google Scholar 

  21. Wang, Y., Qian, W. P., Tan, Y., Ding, S. H., & Zhang, H. Q. (2007). Talanta, 72, 1134–1140.

    Article  CAS  Google Scholar 

  22. Liang, Y., Gong, J. L., & Huang, Y. (2007). Talanta, 72, 443–449.

    Article  CAS  Google Scholar 

  23. Jung, K.-H., Yoon, J.-W., & Koshizaki, N. (2008). Current Applied Physics, 8, 761–765.

    Article  Google Scholar 

  24. Qi, Y. L., Chen, M., Liang, S., & Yang, W. (2008). Applied Surface Science, 254, 1684–1690.

    Article  CAS  Google Scholar 

  25. Fang, L. Y., Lu, Z. Z., Wei, H., & Wang, E. K. (2008). Biosensors & Bioelectronics, 23, 1645–1651.

    Article  CAS  Google Scholar 

  26. Kandimalla, V. B., Tripathi, V. S., & Ju, H. X. (2006). Biomaterials, 27, 1167–1174.

    Article  CAS  Google Scholar 

  27. Xue, J. G., Wang, C. G., & Ma, Z. F. (2007). Materials Chemistry and Physics, 105, 419–425.

    Article  CAS  Google Scholar 

  28. Zhang, L., Feng, Y. G., & Wang, L. Y. (2007). Material Research Bulletin, 42, 1457–1467.

    Article  CAS  Google Scholar 

  29. Miao, Y., & Tan, S. N. (2001). Analytica Chimica Acta, 437, 87–93.

    Article  CAS  Google Scholar 

  30. Yao, H., & Li, N. (2005). Biosensors and Bioelectronics, 21, 372–377.

    Article  CAS  Google Scholar 

  31. Lei, C. X., & Hu, S. Q. (2004). Bioelectrochemistry, 65, 33–39.

    Article  CAS  Google Scholar 

  32. Xiong, H. Y., & Chen, T. (2007). Electrochemistry Communications, 9, 2671–2675.

    Article  CAS  Google Scholar 

  33. Tripathi, V. S., & Kandimalla, V. B. (2006). Biosensors and Bioelectronics, 21, 1529–1535.

    Article  CAS  Google Scholar 

  34. Chen, S. H., Yuan, R., & Chai, Y. Q. (2007). Biosensors and Bioelectronics, 22, 1268–1274.

    Article  CAS  Google Scholar 

  35. Tangkuaram, T., & Ponchio, C. (2007). Biosensors and Bioelectronics, 22, 2071–2078.

    Article  CAS  Google Scholar 

  36. Xiao, Y., & Ju, H. X. (1999). Analytica Chimica Acta, 391, 299–306.

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by the NNSF of China (20675064), the Ministry of Education of China (Project 708073), the Natural Science Foundation of Chongqing City (CSTC-2009BA1003) and High Technology Project Foundation of Southwest University (XSGX02), China.

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Authors and Affiliations

  1. Chongqing Key Laboratory of Analytical Chemistry, College of Chemistry and Chemical Engineering, Southwest University, Chongqing, 400715, People’s Republic of China

    Shirong Yuan, Ruo Yuan, Yaqin Chai, Ying Zhuo, Xia Yang & Yali Yuan

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  1. Shirong Yuan
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  2. Ruo Yuan
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  3. Yaqin Chai
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  4. Ying Zhuo
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  5. Xia Yang
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Correspondence to Ruo Yuan.

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Yuan, S., Yuan, R., Chai, Y. et al. Enzyme biosensor based on the immobilization of HRP on SiO2/BSA/Au composite nanoparticles. Appl Biochem Biotechnol 162, 2189–2196 (2010). https://doi.org/10.1007/s12010-010-8993-1

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