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Applied Biochemistry and Biotechnology

, Volume 174, Issue 3, pp 984–997 | Cite as

Biofunctionalized Gold Nanoparticle-Conducting Polymer Nanocomposite Based Bioelectrode for CRP Detection

  • Sujeet K. Mishra
  • Vikash Sharma
  • Devendra Kumar
  • RajeshEmail author
Article

Abstract

An electrochemical impedance immunosensing method for the detection and quantification of C-reactive protein (αCRP) in phosphate buffered saline (PBS) is demonstrated. The protein antibody, Ab-αCRP, has been covalently immobilized on a platform comprising of electrochemically deposited 3-mercaptopropionic acid-capped gold nanoparticles Au(MPA)-polypyrrole (PPy) nanocomposite film of controlled thickness onto an indium tin oxide-coated glass plate. The free carboxyl groups present on the nanocomposite film have been used to site-specifically immobilize the Ab-αCRP biomolecules through a stable acyl amino ester intermediate generated by N-(3-dimethylaminopropyl)-N′-ethyl carbodiimide hydrochloride and N-hydroxysuccinimide. The nanocomposite film was characterized by atomic force microscopy, high-resolution transmission electron microscopy, Fourier transform infrared spectroscopy, and electrochemical techniques. The bioelectrode was electrochemically analyzed using modified Randles circuit in terms of constant phase element (CPE), electron transfer resistance (R et), and Warburg impedance (Z w). The value of n, a CPE exponent used as a gauge of heterogeneity, for the Au-PPy nanocomposite film was found to be 0.56 which is indicative of a rather rough morphology and porous structure. A linear relationship between the increased ∆R et values and the logarithmic value of protein antigen, Ag-αCRP, concentrations was found in the range of 10 ng to 10 μg mL−1 with a R et sensitivity of 46.27 Ω cm2/decade of [Ag-αCRP] in PBS (pH 7.4).

Keywords

Electrochemical impedance spectroscopy Conducting polymer Gold nanoparticles Protein immobilization 

Notes

Acknowledgments

We are grateful to Prof. R. C. Budhani, Director, National Physical Laboratory, New Delhi, India for providing the facilities. S.K. Mishra is thankful to the Council of Scientific and Industrial Research, India for providing a senior research fellowship (SRF). We also acknowledge Mr. Sandeep Singh for AFM measurements.

References

  1. 1.
    Ates, M. (2013). Materials Science and Engineering: C, 33, 1853–1859.CrossRefGoogle Scholar
  2. 2.
    Yuan, L., Wei, W., & Liu, S. (2012). Biosensors and Bioelectronics, 38, 79–85.CrossRefGoogle Scholar
  3. 3.
    Revin, S. B., & John, S. A. (2012). Analyst, 137, 209–215.CrossRefGoogle Scholar
  4. 4.
    Park, S. J., Kwon, O. S., & Jan, J. (2013). Chemical Communications, 49, 4673–4675.CrossRefGoogle Scholar
  5. 5.
    Singh, M., Kathuroju, P. K., & Jampana, N. (2009). Sensors and Actuators, B, 143, 430–443.CrossRefGoogle Scholar
  6. 6.
    Jia, Y., Xiao, P., He, H., Yao, J., Liu, F., Wang, Z., et al. (2012). Applied Surface Science, 258, 6627–6631.CrossRefGoogle Scholar
  7. 7.
    Dong, Z. H., Wei, Y. L., Shi, W., & Zhang, G. A. (2011). Materials Chemistry and Physics, 131, 529–534.CrossRefGoogle Scholar
  8. 8.
    Shi, W., & Ma, Z. (2011). Biosensors and Bioelectronics, 26, 3068–3071.CrossRefGoogle Scholar
  9. 9.
    Cao, X., Ye, Y., & Liu, S. (2011). Analytical Biochemistry, 417, 1–16.CrossRefGoogle Scholar
  10. 10.
    Rajesh, Sharma, V., Mishra, S. K., & Biradar, A. M. (2012). Materials Chemistry and Physics, 132, 22–28.CrossRefGoogle Scholar
  11. 11.
    Qiu, L., Peng, Y., Liu, B., Lin, B., Peng, Y., Malik, M. J., et al. (2012). Applied Catalysis A, 413, 230–237.CrossRefGoogle Scholar
  12. 12.
    Ledue, T., & Rifai, N. (2003). Clinical Chemistry, 49, 1258–1271.CrossRefGoogle Scholar
  13. 13.
    Kindt, T., Osborne, B., & Golsby, R. (2007). Kuby immunology (6th ed.). New York: W.H. Freeman.Google Scholar
  14. 14.
    Algarra, M., Gomes, D., & Esteves da Silva, J. C. G. (2012). Clinica Chimica Acta, 415, 1–9.CrossRefGoogle Scholar
  15. 15.
    McDonnell, B., Hearty, S., Leonard, P., & O′ Kennedy, R. (2009). Clinical Biochemistry, 42, 549–561.CrossRefGoogle Scholar
  16. 16.
    Lin, K. C., Kunduru, V., Bothara, M., Rege, K., Prasad, S., & Ramakrishna, B. L. (2010). Biosensors and Bioelectronics, 25, 2336–2342.CrossRefGoogle Scholar
  17. 17.
    Yang, T., Wang, S., Jin, H., Bao, W., Huang, S., & Wang, J. (2013). Sensors and Actuators, B, 178, 310–315.CrossRefGoogle Scholar
  18. 18.
    Suprun, E. V., Shilovskaya, A. L., Lisitsa, A. V., Bulko, T. V., Shumyantseva, V. V., & Archakov, A. I. (2011). Electroanalysis, 23, 1051–1057.CrossRefGoogle Scholar
  19. 19.
    Xu, H., Wang, L., Ye, H., Yu, L., Zhu, X., Lin, Z., et al. (2012). Chemical Communications, 48, 6390–6392.CrossRefGoogle Scholar
  20. 20.
    Guo, X., Kulkarni, A., Doepke, A., Halsall, B. H., Iyer, S., & Heineman, W. R. (2012). Analytical Chemistry, 84, 241–246.CrossRefGoogle Scholar
  21. 21.
    Wei, Q., Zhao, Y., Du, B., Wu, D., Li, H., & Yang, M. (2012). Food Chemistry, 134, 1601–1606.CrossRefGoogle Scholar
  22. 22.
    Shi, Y., Wang, H., & Cai, N. (2012). Journal of Power Sources, 208, 24–34.CrossRefGoogle Scholar
  23. 23.
    Sen, T., & Patra, A. (2009). Journal of Physical Chemistry C, 113, 13125–13132.CrossRefGoogle Scholar
  24. 24.
    Shamsipur, M., Asgari, M., Maragheh, M. G., & Moosavi-Movahedi, A. A. (2012). Bioelectrochemistry, 83, 31–37.CrossRefGoogle Scholar
  25. 25.
    Davies, T. J., Banks, C. E., & Compton, R. G. (2005). Journal of Solid State Electrochemistry, 9, 797–808.CrossRefGoogle Scholar
  26. 26.
    Compton, R. G., & Banks, C. E. (2007). Understanding voltammetry. London: World Scientific.CrossRefGoogle Scholar
  27. 27.
    Solomon, A., Mamuru, Kenneth, I., & Ozoemena, I. (2010). Electroanalysis, 22, 985–994.CrossRefGoogle Scholar
  28. 28.
    Bryan, T., Luo, X., Bueno, P. R., & Davis, J. J. (2013). Biosensors and Bioelectronics, 39, 94–98.CrossRefGoogle Scholar
  29. 29.
    Zhu, Jie, J., Xu, Zhong, J., He, Tao, J., et al. (2003). Analytical Letters, 36, 1547–1556.CrossRefGoogle Scholar
  30. 30.
    Quershi, A., Gurbuz, Y., Kang, W. P., & Davidson, J. L. (2009). Biosensors and Bioelectronics, 25, 877–882.CrossRefGoogle Scholar
  31. 31.
    Zhou, F., Lu, M., Wang, W., Bian, Z. P., Zhang, J. R., & Zhu, J. J. (2010). Clinical Chemistry, 56, 1701–1707.CrossRefGoogle Scholar
  32. 32.
    Puri, N., Tanwar, V. K., Sharma, V., Ahuja, T., Biradar, A. M., & Rajesh. (2010). International Journal of Integrative Biology, 9, 1–5.Google Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Sujeet K. Mishra
    • 1
    • 2
  • Vikash Sharma
    • 1
  • Devendra Kumar
    • 2
  • Rajesh
    • 1
    Email author
  1. 1.CSIR-National Physical LaboratoryNew DelhiIndia
  2. 2.Department of Applied ChemistryDelhi Technological UniversityDelhiIndia

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