항원인 마뇨산에 결합된 도파민을 이용한 전기화학적 면역 분석법
Electrochemical Immunoassay based on the Dopamine-antigen Conjugate for Detecting Hippuric Acid
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초록
이 논문에서는 체내에 존재하는 작은 유기물의 하나인 마뇨산을 빠르고 정량적으로 검출하기 위하여 전기화학적 면역분석법을 이용하였다. 마뇨산은 톨루엔이 인체에 노출되었을 때의 주된 신진 대사 물질로서 대표적인 생화학적 지표물질이다. 톨루엔 노출에 대한 신속하고 정확한 관찰은 산업현장에서 건강관리를 위하여 매우 중요하다. 그래서 마뇨산을 빠르고 정확하게 검출하기 위하여 전기화학적 면역분석법을 실시하였다. 본 연구에서 제시하는 새로운 면역분석방법은 전기화학적 활성물질의 하나인 도파민을 측정물질인 마뇨산과 직접 공유결합을 통하여 항원이 포함된 복합체를 합성하였다. 전기화학적 측면에서 도파민은 두 개의 하이드록시기를 가지고 있어 매우 뛰어난 산화/환원신호를 보인다. 또한 도파민-마뇨산 복합물도 뛰어난 산화/환원 신호를 보이기 때문에 면역 분석에적합하다. 도파민-마뇨산 복합물과 마뇨산의 항체와의 균질경쟁반응을 전기화학적인 방법을 통하여관찰하였다. 본 면역분석을 통하여 실시한 결과는 마뇨산의 농도가 0.010~2.500 mg/mL 까지 정량적으로 분석됨으로써 실제 면역 센서에 적용할 수 있음을 확인하였다.
Abstract
In this work, we describe an electrochemical immunosensor for simple, fast andquantitative detection of a urinary hippuric acid (HA). Urinary HA, of molecular weight 180DA, is one of the major metabolites and biological indicators in toluene-exposed humans. Simpleand ubiquitous monitoring of exposure to toluene is very important in occupational healthcare. We propose the electrochemical immunoassay based on the dopamine-antigen conjugatefor detecting hippuric acid. Our electrochemical immunoassay system employs a conjugate ofdopamine (DA) as an electrochemical active molecule and hippuric acid (HA) as an antigen. As an electrochemical aspect, dopamine (DA) containing two hydroxyl group can show excellentredox signal. Also, dopamine-tethered hippuric acid (DA-HA) shows the reversible redoxsignal in the immunoassay. The competition between HA and DA-HA generated electric signals proportional to HA concentration. The electrochemical immunoassay was performed with DAHAon the screen printed carbon electrodes (SPCEs), and then applies the mixture antigen (HA)and HA-antibody. The electrical signals were proportional to HA in the range of 0.010~2.500 mg/mL which is enough range to be used for the point-of-care.
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