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바이오나노 입자의 제조 및 시그널 증폭 프로브로의 응용

Title
바이오나노 입자의 제조 및 시그널 증폭 프로브로의 응용
Other Titles
Fabrication of Bio-Nano Particle and Its Application as Signal Amplification Probe
Author
김남현
Alternative Author(s)
Kim, Nam-Hyun
Advisor(s)
성기훈
Issue Date
2007-02
Publisher
한양대학교
Degree
Master
Abstract
이 연구의 목적은 미세질량저울 (Quartz Crystal Microbalance, QCM) 에서 매우 낮은 농도의 타겟 분자들을 고감도로 검출하기 위하여 금 나노 입자를 시그널 증폭 프로브로 사용하는 것이다. Streptavidin-biotin 상호작용을 기본 모델로 하여 우선 biotin이 수식된 금 나노 입자가 오랜 시간 안정된 상태로 흩어져 있도록 biotin의 농도를 조절하였다. 금 나노 입자의 표면은 UV-visspectra와 AFM image로 분석하였다. 그 결과 10 mM의 Biotin-HPDP를 사용하였을때, 금 나노 입자의 엉김 현상이 발생하지 않으므로 이 시스템에 사용하기 적합했다. QCM에서 타겟분자인 streptavidin을 검출하기 위해서 QCM 전극의 금 표면에 biotin으로 수식된 BSA(bovine serum albumin)를 흡착시켰다. 즉, 표면의 Biotin과 타겟분자인 streptavidin을 coupling시켜 sensing interface를 만들어 주는 것이다. Sensing 표면에 target streptavidin과 10mM Biotin-HPDP가 modify된 금 나노 입자를 반응시킨다. Biotinylated 금 나노 입자는 signal증폭 probe로 사용하는데 검출한계를 50 ng/ml까지 향상 시켰다. Streptavidin의 detection system에서 signal target molecule인 streptavidin의 농도가 1 ng/ml~10 ml/ml인 넓은 범위로 frequency change를 분석한 결과 매우 좋은 직선형을 보였다. 또한 dissipation 변화를 측정한 결과 금 전극위의 biotin-BSA의 흡착 layer와 Biotin-BSA에 streptavidin이 assembly된 layer는 매우 compact하고 rigid하다 반면에 streptavidin layer 위의 Biotinylated gold 나노 입자의 구조는 flexible하고 dissipative한 모습으로 sensing 표면에서 가늘고 길게 뻗어 있는 구조를 하고 있다.
We report here a novel strategy for the high-sensitive detection of target biomolecules with very low concentrations on a quartz crystal microbalance (QCM) device using gold nanoparticles as signal enhancement probes. By employing a streptavidin-biotin interaction as a model system, we could prepare biotin-conjugated gold nanoparticles maintaining good dispersion and long-term stability by controlling the biotin density on the surface of gold nanoparticles that have been investigated by UV-vis spectra and AFM images. These results showed that 10 mM N-(6-[biotinamido]hexyl)-3'-(2'-pyridyldithio)propionamide (biotin-HPDP) was the critical concentration to prevent the nonspecific aggregation of gold nanoparticles in this system. For sensing streptavidin target molecules by QCM, biotinylated BSA was absorbed on the Au surface of the QCM electrode and subsequent coupling of the target streptavidin to the biotin in the sensing interface followed. Amplification of the sensing process was performed by the interaction of the target streptavidin on the sensing surface with gold nanoparticles modified with 10 mM biotin-HPDP. The biotinylated gold nanoparticles were used as signal amplification probes to improve the detection limit, which was 50 ng/ml, of the streptavidin detection system without signal enhancement, and the calibration curve determined for the net frequency changes showed good linearity over a wide range from 1 ng/ml to 10 mg/ml for the quantitative streptavidin target molecule analysis. In addition, the measured dissipation changes suggested that the layer of biotin-BSA adsorbed on the Au electrode and the streptavidin layer assembled on the biotin-BSA surface were highly compact and rigid. On the other hand, the structure formed by the biotinylated gold nanoparticles on the streptavidin layer was flexible and dissipative, being elongated outward from the sensing surface.
URI
http://dcollection.hanyang.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000048818https://repository.hanyang.ac.kr/handle/20.500.11754/150633
Appears in Collections:
GRADUATE SCHOOL[S](대학원) > BIONANOTECHNOLOGY(바이오나노학과) > Theses (Master)
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