研究了纳米颗粒的多酶活性,包括过氧化物酶和过氧化氢酶,以及在葡萄糖高灵敏检测中的应用.
Zhang et al.0.18
Prussian Blue Modi ed Ferritin as Peroxidase Mimetics and Its Applications in Biological Detection
4. CONCLUSIONSIn summary, a very simple and cheap template method to obtain PB-Ft NPs with very small size has been developed and the combining functions of the mimetic enzyme activity from PBNPs and the speci city of ferritin were realized. It has been demonstrated that the peroxidaselike activity of PB-Ft NPs is pH, temperature and PB-Ft NPs concentration dependent and ts well with typical Michaelis–Menten kinetics. PB-Ft NPs displayed quite high sensitivity and af nity to H2 O2 with ABTS as choromogenic substrate, which makes PB-Ft NPs a good reagent in glucose detection. PB-Ft NPs were successfully used in the ELISA, indicating that in PB-Ft NPs the antibody binding functions of ferritin were preserved. These studies show this composite nanoparticle has potential application potential in bio-detection. Acknowledgments: This research was supported by the National Important Science Research Program of China (Nos. 2011CB933503, 2013CB733800), National Natural Science Foundation of China (Nos. 31170959, 30970787), the Basic Research Program of Jiangsu Province (Natural Science Foundation, Nos. BK2011036, BK2009013), Research Fund for the Doctoral Program of Higher Education of China (20110092110029).0 10 20 30 40 50 60 70
Absorbance (415 nm)
0.16Absorbance (415 nm)
0.14 0.12 0.10 0.08 0.06 0 100
0.05 0.04 0.03 0.02 0.01 0.00 0 1 2 3 4 5 6 7 y=0.00779x+0.00328 R=0.99423 range: 0.39µM~ 6.25µM
Concentration of glucose (µM)
200
300
400
Concentration of glucose (µM)Fig. 7. A dose-response curve for glucose detection using GOx and PBFt NPs. Inset: linear calibration plot for glucose. The error bars represent the standard deviation of three measurements.
(a) Absorbance (650 nm)
1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4
PB-Ft concentration (10–11M) (b)Absorbance (650 nm)1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0 control-1 control-2 competition 1µg/mL 2.5µg/mL 5µg/mL 10µg/mL
References a
nd Notes
RESEARCH ARTICLE
Fig. 8. Enzyme-linked immunosorbent assay based on both the peroxidase-like activity and the speci city of PB-Ft NPs. (a) Effect of PBFt NPs concentration on the detection of anti-HoSF (b) Immunoassays for the anti-HoSF antibody of different concentrations. The control groups using PBS (control-1) and HoSF (control-2) instead of PB-Ft NPs as well as the HoSF competition group were also carried out for comparison.
a inevitable catalytic reaction. PB-Ft NPs group shows a much higher absorbance which increases with the concentration of anti-HoSF antibody coated in the well, indicating PB-Ft NPs can speci cally bind to the anti-HoSF antibody. Very currently, Yan’s group19 demonstrated that magnetoferritin (M-HFn) nanoparticles synthesized by encapsulating iron oxide nanoparticles inside a HFn shell can target the overexpressed transferrin receptor 1 (TfR1) in tumour cells. It is expected that PB-Ft NPs, compared to Yan’s work, may have higher ef ciency due to the higher peroxidase-like activity of PBNPs than iron oxide nanoparticles.J. Nanosci. Nanotechnol. 12, 1–8, 2012
1. L. Z. Gao, J. Zhuang, L. Nie, J. B. Zhang, Y. Zhang, N. Gu, T. H. Wang, J. Feng, D. L. Yang, S. Perrett, and X. Y. Yan, Nature Nanotech. 2, 577 (2007). 2. H. D. Hill, R. A. Vega and C. A. Mirkin, Anal. Chem. 79, 9218 (2007). 3. S. Boland, F. Barriere, and D. Leech, Langmuir 24, 6351 (2008). 4. M. Nag, A. Patel, and N. M. Rao, J. Biomed. Nanotechnol. 7, 42 (2011). 5. M. M. Rahman, J. Biomed. Nanotechnol. 7, 351 (2011). 6. L. J. Xu, J. J. Du, Y. Deng, and N. Y. He, J. Biomed. Nanotechnol. 8, 1006 (2012). 7. F. Wang, C. Ma, X. Z., C. Y. Li, Y. Deng, and N. Y. He, J. Biomed. Nanotechnol. 8, 786 (2012). 8. J. Li, J. D. Qiu, J. J. Xu, H. Y. Chen, and X. H. Xia, Adv. Funct. Mater. 17, 1574 (2007). 9. X. Q. Zhang, S. W. Y. Gong, Y. Zhang, T. Yang, C. Y. Wang, and N. Gu, J. Mater. Chem. 20, 5110 (2010). 10. A. A. Karyakin, E. E. Karyakina, and L. Gorton, Anal. Chem. 72, 1720 (2000). 11. D. Ellis, M. Eckhoff, and V. D. Neff, J. Phys. Chem. 85, 1225 (1981). 12. G. C. Ford, P. M. Harrison, D. W. Rige, J. M. Smith, A. Treffry, J. L. White, and J. Yarriv, Phil. Trans. R Soc. Lond. 304, 551 (1984). 13. D. M. Lawson, P. J. Artymiuk, S. J. Yewdall, J. M. A. Smith, J. C. Livingstone, A. Treffry, A. Luzzzago, S. Levi, P. Arosio, G. Cesareni, C. D. Thomas, W. V. Shaw, and P. M. Harrison, Nature 349, 541 (1991). 14. M. V. Darl, P. M. Harrison, and W. Bottke, Eur. J. Biochem. 222, 367 (1994). 15. P. M. Harrison and P. Arosio, Biochimica et Biophysica Acta 1275, 161 (1996). 16. C. Q. Cao, L. X. Tian, Q. S. Liu, W. F. Liu, G. J. Chen, and Y. X. Pan, J. Geophys. Res. 115, B07103 (2010).