Development of ultrasensitive biological immunoassay system using Fe3O4 nanoparticles and magnetic sensor in the liquid phase

Paper Details

Research Paper 01/03/2017
Views (205) Download (3)

Development of ultrasensitive biological immunoassay system using Fe3O4 nanoparticles and magnetic sensor in the liquid phase

Md. Anwarul Kabir Bhuiya, Raihana Ferdaws, Masaki Asai, Yuichi Higuchi, Takeshi Yoshida, Keiji Enpuku, Edmund Soji Otabe
Int. J. Biosci.10( 3), 406-417, March 2017.
Certificate: IJB 2017 [Generate Certificate]


In this paper, we first describe details of the measurement system (MR and Flux Gate sensor). The measurement system using the MR sensor showed a sensitivity to detect 1.4 × 107 of the markers in 60 µl of solution. The sensitivity was improved as 8.3 × 106 when the flux gate sensor was used.  The sensitivity of the present method was estimated as 3.8 × 10-16 and 2.3×10-16 mol/ml in terms of the molecular-number concentration for the MR and the flux gate sensor, respectively. We next demonstrate the detection of biological targets known as biotins, which were conjugated on the surface of the polystyrene beads with a diameter of 3.3 µm. The minimum detectable number of beads was Np = 10,000 and 5,000 for the case of the MR and the flux gate sensor, respectively. Since about 700 biotins were fixed on the single polymer bead, the minimum detectable number of biotins was estimated as Nb = 7 × 106 and 3.5× 106 for the case of the MR and the flux gate senor, respectively.  A strong relationship was obtained between the number of bound markers and the number of biotin-conjugated polymer beads, which confirmed the validity of the method. The detection sensitivity can be estimated as 1.9× 10-16 and 0.8 × 10-16 mol/ml in terms of the molecular-number concentration of biotin for the MR and the flux gate sensor, respectively. These results are consistent with the estimated sensitivity of the measurement system.


  1.  Li, S. Sun, R. J. Wilson, R. L. White, Nader Pourmand and S. X.Wang, Sens. Actuators A 126, 98 (2006).
  2. Li, S. X. Wang and S. Sun, IEEE Trans. Magn. 40, 3000 (2004).
  3. X. Wang, S.-Y. Bae, G. Li, S. Sun, R. L. White, J. T. Kemp and C. D. Webb, J. Magn. Magn. Mater. 293 731.
  4. Manual of Magnetoresistive and Fluxgate sensor.
  5. Schotter, P. B. Kamp, A. Becker, A. Puhler, D. Brinkmann, W. Schepper, H. Bruckl and G. Reiss, IEEE Trans. Magn. 38, 3365 (2002).
  6. Magnetic Sensors and magnetometers (P. Ripka et al) ISBN: 1580530575.
  7. Payet, D. Vincent, L. Delaunary, and G. Noyel: J. Magn. Magn. Mater. 186 (1998) 168.
  8. Enpuku, T. Tanaka, Y. Tamai, F. Dang, N. Enomoto, J. Hojo, H. Kanzaki, and N. Usuki: Jpn. J. Appl. Phys. 47 (2008) 7859.
  9. X. Wang and G. Li, “Advances in giant magneto resistance biosensors with magnetic nanoparticle tags: Review and outlook,” IEEE Trans. Magn., vol. 44, pp. 1687–1702, 2008.
  10. B. Hong, H. J. Krause, K. B. Song, C. J. Choi, M. A. Chung, S. W. Son, and A. Offenhausser, “Detection of two different influenza A viruses using a nitrocellulose membrane and a magnetic biosensor,” J. Immunol. Meth., vol. 365, pp. 95–100, 2011.
  11. Gaster, L.Xu, S. J. Han, R. J. Wilson,D.A. Hall, S. J.Osterfeld, H. Yu, andS.X.Wang, “Quantification of protein interactions and solution transport using high-density GMR sensor arrays,” Nat. Nanotechnol., vol. 6, pp. 314–320, 2011.
  12. Grossman, W. Myers, V. Vreeland, R. Bruehl, M. D. Alper, C. R. Bertozzi, and J. Clarke, “Determination of bacteria in suspension using a superconducting quantum interference device,” PNAS U. S. A., vol. 101, pp. 129–134, 2004.
  13. Fornara, P. Johansson, K. Petersson, S. Gustafsson, J. Qin, E. Olsson, D. Ilver, A. Krozer, M. Muhammad, and C. Johansson, “Tailored magnetic nanoparticles for direct and sensitive detection of biomolecules in biological samples,” Nano Lett., vol. 8, pp. 3423–3428, 2008.
  14. Stromberg, T. Torre, J. Goransson, K. Gunnarsson, M. Nilson, P. Svedlindh, and M. Stromme, “Multiplex detection of DNA sequences using the volume-amplified magnetic nanobead detection assay,” Anal. Chem, vol. 81, pp. 3398–3406, 2009.
  15. Enpuku, Y. Tamai, T. Mitake, T. Yoshida, andM.Matsuo, “AC susceptibility measurement of magnetic markers in suspension for liquid phase immunoassay,” J. Appl. Phys., vol. 108, p. 034701, 2010.
  16. J.Chiu, H.E. Horng, J. J. Chien, S.H. Liao,C.H.Chen, B. Y. Shih, C. C. Yang, C. L. Lee, T. F. Chen, S. Y. Yang, C. Y. Hong, and H. C.
  17. Yang, “Multi-channel SQUID-based ultra-high-sensitivity in-vitro detections for bio-markers of Alzheimer’s disease via immunomagnetic reduction,” IEEE Trans. Appl. Supercond., vol. 21, pp. 477–480, 2011.
  18. K. Bhuiya, T. Mitake, M. Asai, T. Ito, S. Chosakabe, T. Yoshida, K. Enpuku, and A. Kandori, “Liquid-phase immunoassays using Brownian relaxation of magnetic markers,” IEEE Trans. Magn., vol. 47, pp. 2867–2870, 2011.
  19. Enpuku, H. Watanabe, Y. Higuchi, T. Yoshida, H. Kuma, N. Hamasaki, M. Mitsunaga, H. Kanzaki, and A. Kandori, “Characterization of magnetic markers for liquid-phase immunoassays usingBrownian relaxation,” Jpn. J. Appl. Phys., vol. 51, p. 023002, 2012.
  20. Anwarul Kabir BHUIYA, Masaki ASAI, Takashi YOSHIDA and Keiji ENPUKU, “Magnetic Sensor Based Liquid-Phase Immunoassays for the Detection of Biological Targets”. Research reports on information science and electrical engineering of Kyushu University 16(2) p45-50; 2011-09-26.