Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Detection of Carp Vitellogenin with Piezoelectric Immunosensor

압전류적 면역센서에 의한 잉어 Vitellogenin 검출

  • Published : 2006.09.30
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Abstract

Vitellogenin has been known as a potent biomarker protein for the estrogenic activity in fish exposed to endocrine disruptors. In this study, a piezoelectric immunosensor making use of an anticarp vitellogenin antibody and an AT-cut quartz crystal microbalance as the biological component and transducer was prepared, followed by its application to the analysis of carp vitellogenin as follows. Antibody immobilization was conducted by chemisorption of a thiolated antibody with a heterobifunctional thiolation cross-linker, sulfosuccinimidyl 6-[3-(2-pyridyldithio)propionamido]hexanoate. The reaction buffer for the immunosensor system was optimized as 0.1 M sodium phosphate (pH 7.4). Concentration-dependent sensor responses were obtained in the vitellogenin concentrations ranging from 0.4864 to 486.4000 nM, with a linear correlation between vitellogenin concentration and frequency shift in double-logarithmic scale. The limit of detection of the immunosensor for carp vitellogenin was presumed as 0.4864 nM.

내분비계장애물질에 노출된 어류의 에스트로겐 활성에 대한 생물지표단백질인 vitellogenin을 검출하는 압전류적 면역센서를 잉어 vitellogenin에 대한 항체와 AT-cut 수정진동자를 생물요소와 변환기로 사용하여 구성하고 이를 이용한 잉어 vitellogenin 검출을 행하였다. 이형이기능성의 티올화 가교화제인 sulfosuccinimidyl 6-[3-(2-pyridyldithio)propionamido]hexanoate 처리에 의하여 티올화된 항체를 수정진동자상의 금전극에 화학흡착법에 의하여 고정화하여 센서 chip을 제조하였다. 센서반응을 위한 반응완충용액을 0.1M sodium phosphate(pH 7.4)로 선정한 후 $0.4864{\sim}486.4000\;nM$의 vitellogenin 용액을 가하였을 때 농도의존적인 센서반응의 증가가 나타났으며 이 때 잉어 vitellogenin에 대한 검출한계는 0.4864 nM로 추정되었다.

Keywords

References

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