On-chip type cation-exchange chromatography with ferrocene-labeled anti-hemoglobin antibody and electrochemical detector for determination of hemoglobin A1c level

Tsuyoshi Tanaka, Kojiro Izawa, Mina Okochi, Tae Kyu Lim, Shugo Watanabe, Manabu Harada, Tadashi Matsunaga

Research output: Contribution to journalArticlepeer-review

23 Citations (Scopus)

Abstract

An on-chip type cation-exchange chromatography system with electrochemical detection of HbA1c, which is one of the most important diabetes marker protein, was developed using ferrocene-conjugated anti-human hemoglobin (Hb) monoclonal antibody (FcAb). The FcAb was used as an electrochemical probe for the detection of each Hb. The system contains syringe pump, on-chip type cation-exchange column consisted of PDMS and cation-exchange resin beads, and a three-electrode flow-cell system. The separation conditions of HbA1c in blood calibrator samples from other Hbs, e.g. HbA0, HbA1a or HbA1b, were optimized using the on-chip type system. The electrochemical oxidation current from FcAb reacting with each Hb was measured at 350 mV (vs. Ag/AgCl). Hbs including HbA1a and HbA1b, HbA1c and HbA0 fractions were eluted in this order. A linear relationship between HbA1c levels and electrochemical oxidation currents was obtained in the range from 4.0% to 12.6% HbA1c. All procedure including antigen-antibody reaction was completed in 15 min. Furthermore, a good correlation was obtained between KO500 method (HPLC) and our proposed method. These results indicate that the on-chip type system with electrochemical detection can be applied to a novel POCT device for rapid and precise detection of HbA1c.

Original languageEnglish
Pages (from-to)186-190
Number of pages5
JournalAnalytica Chimica Acta
Volume638
Issue number2
DOIs
Publication statusPublished - 2009 Apr 13
Externally publishedYes

Keywords

  • Electrochemical detection
  • Ferrocene-conjugated antibody
  • Hemoglobin A
  • On-chip type cation-exchange chromatography system

ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry
  • Spectroscopy
  • Environmental Chemistry

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