Fish Physiology and Biochemistry

, Volume 39, Issue 2, pp 299–308 | Cite as

Carbon nanotube enhanced label-free immunosensor for amperometric determination of oocyte maturation-inducing hormone in fish

  • Mutsuko Hirai
  • Tadayoshi Muramatsu
  • Hitoshi Ohnuki
  • Kyoko Hibi
  • Huifeng Ren
  • Hideaki Endo


Maintaining high-quality fish eggs stably and efficiently is important for aquaculture. We developed a label-free immunosensor system for measuring 17,20β-dihydroxy-4-pregnen-3-one (DHP). DHP is suddenly secreted before ovulation as a maturation-inducing hormone in fish, and therefore, DHP levels are an indicator for predicting ovulation. The method is based on immunologic reactions and amperometric measurement using cyclic voltammetry (CV). For biomolecular immobilization on the surface of sensing electrode, Au electrode, we used self-assembled monolayers of thiol-containing compounds to fix anti-DHP immunoglobulin. In addition, we used a single-walled carbon nanotube to improve sensitivity. Using this electrode, we were able to determine the CV signal change caused by the antigen–antibody complex. The proposed immunosensor system showed a linear correlation (correlation coefficient: 0.9827) between the anodic peak current of the CV and the DHP level in range from 15.6 to 50,000 pg ml−1. The sensor system was then applied to monitor DHP of goldfish (Carassius auratus). Blood plasma of fish was collected every 3 h after administering a DHP inducer. In the measurement, the anodic peak current of the CV showed distinct changes depending on DHP levels in the blood plasma. A good relationship was observed between DHP levels determined by our proposed system and the conventional method (correlation coefficient: 0.9351).


17,20β-Dihydroxy-4-pregnen-3-one Ovulation Biosensor Carbon nanotube Immunoassay Label-free Fish 



This research was supported in part by a Grant-in-Aid for Scientific Research (B) from The Ministry of Education, Culture, Sports, Science, and Technology. We wish to thank Mr. Junzo Yana (Institute of Carbon Science and Technology, Shinshu University) for their helpful discussions.


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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Mutsuko Hirai
    • 1
  • Tadayoshi Muramatsu
    • 1
  • Hitoshi Ohnuki
    • 2
  • Kyoko Hibi
    • 1
  • Huifeng Ren
    • 1
  • Hideaki Endo
    • 1
  1. 1.Faculty of Marine ScienceTokyo University of Marine Science and TechnologyMinato-ku, TokyoJapan
  2. 2.Faculty of Marine TechnologyTokyo University of Marine Science and TechnologyKoto-ku, TokyoJapan

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