• [Communication]
  • Synthesis of Enzyme-Containing PEG Hydrogel Nanospheres for Optical Biosensors
  • Kim B
  • 광바이오센서용 효소를 함유한 PEG 수화젤 나노입자의 합성
  • 김범상
Abstract
In this work, as the first step to develop optical biosensors for a single cell level analysis, the preparation method of nano-scale polymer hydrogel spheres containing an enzyme was set up and the feasibility of the spheres as optical biosensors was investigated. The horseradish peroxidase (HRP) was encapsulated in the PEG hydrogel spheres by suspension photopolymerization, yielding spheres of the average size of 305 nm. After the polymerization, the incorporation and activity of HRP within the spheres were determined by the production of fluorescence resulted from the enzymatic reaction between HRP and H2O2. The fluorescence emission response of the HRP-loaded PEG hydrogel spheres increased by nearly 300% as hydrogen peroxide concentration was changed from 0 to 11 nM in the presence of Amplex Red. The results suggest that the method to prepare the PEG hydrogel nanospheres containing an enzyme could be used for developing optical biosensors to measure various analytes in the very small samples like a single cell.

본 연구에서는 한 개의 세포와 같은 미세한 생체시료 내부의 분석 대상물질을 감지하는데 사용할 수 있는 광바이오센서를 개발하기 위한 첫단계로서 효소가 함유된 고분자 수화젤 구형입자를 나노크기로 중합하는 방법을 확립하고 센서로서의 사용 가능성을 확인하였다. 현탁 광중합을 통하여 305 nm의 평균크기를 가지는 horseradish peroxidase(HRP)가 함유된 poly(ethylene glycol)(PEG) 수화젤 구형입자를 합성하였으며, 중합반응 이후 입자내부의 효소의 존재 및 활성유지를 HRP와 H2O2의 효소반응에 의한 Amplex Red의 형광산화물 생성을 통하여 확인하였다. 합성된 HRP가 함유된 PEG 수화젤 입자는 Amplex Red의 존재하에 H2O2의 농도가 0에서 11 nM로 미량 변화함에 따라서 형광세기가 약 300% 증가함을 보여 주었다. 이러한 결과는 효소가 함유된 PEG 수화젤 나노입자를 합성하는 본 기술이 향후 미세한 생체시료 내부의 다양한 분석대상물질을 감지할 수 있는 나노크기의 광바이오센서를 개발하는데 이용 될 수 있는 가능성을 보여준다.

Keywords: enzyme encapsulation; optical biosensor; PEG hydrogel; nanosphere; hydrogen peroxide

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    ISSN 2234-8077(Online)
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This Article

  • 2005; 29(6): 613-616

    Published online Nov 25, 2005

  • Received on Jun 29, 2005
  • Accepted on Sep 29, 2005