A competitive immunochromatographic assay for testosterone based on electrochemical detection

Kumi Inoue, Pascal Ferrante, Yu Hirano, Tomoyuki Yasukawa, Hitoshi Shiku, Tomokazu Matsue

Research output: Contribution to journalArticlepeer-review

45 Citations (Scopus)


An immunochromatographic assay using nitrocellulose membrane was combined with electrochemical detection using an electrode chip in order to quantitatively detect testosterone as a model analyte. The electrode chip consisted of a gold working electrode, a counter electrode and a pseudo-reference electrode, all fabricated on the bottom of a 3.2 mm × 3.2 mm well. Competitive immunoreactions on the membrane were initiated by flowing a solution containing testosterone and horseradish peroxidase (HRP)-labeled testosterone (a competitor) over the membrane. Prepared membrane was placed in a solution containing ferrocenemethanol (FcOH) and H2O2 in the well of the electrode chip, and the enzyme reaction was detected by amperometry. Labeled HRP captured on the membrane catalyzed the oxidation of FcOH to the oxidized form FcOH+, which was reduced electrochemically by the electrode chip. The electrochemical response of the reduction current decreased with increasing concentration of testosterone over the range 1-625 ng/ml.

Original languageEnglish
Pages (from-to)886-892
Number of pages7
Issue number5
Publication statusPublished - 2007 Oct 31


  • Amperometry
  • Competitive-ELISA
  • Immunochromatography
  • Nitrocellulose membrane
  • Testosterone

ASJC Scopus subject areas

  • Analytical Chemistry
  • Chemistry(all)
  • Biochemistry
  • Spectroscopy


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