• 제목/요약/키워드: Surface plasmon resonance(SPR)

검색결과 126건 처리시간 0.023초

SPR 근거 DNA 칩을 이용한 페놀 화합물 특이 CapR 조절 단백질과 촉진유전자와의 상호작용 연구 (Interaction of Phenolic Compound-Specific Activator with Its Promoter using SPR-Based DNA Chip)

  • 박선미;박후휘;임운기;신혜자
    • 생명과학회지
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    • 제13권1호
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    • pp.99-104
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    • 2003
  • Aromatic compounds are of major concern among environmental pollutants due to their toxicity and persistence. To monitor aromatic compounds in a real time with a better sensitivity, a new method of SPR (surface plasmon resonance) based on DNA chip (Biacore 3000) was developed here. It is thought that CapR regulatory protein as a complex with phenol, could bind to their corresponding promoter, Po. Biotinylated DNA oligomers for the promoter was synthesized by PCR and coupled onto streptoavidin-linked CM5-chip. CapR regulatory proteins were purified after cloning their genes in pET21a (+) vector and expressing proteins. The interaction was assessed by the system where the regulatory proteins flowed with or without phenol through the cells of DNA chip. CapR regulatory protein even in the presence of phenol had no response to its promoter, Po, suggesting that other factor(s) might be required for the activation of Po promoter. The present work reveals a promising possibility of the SPR-based DNA chip in monitoring specific environmental pollutants in a real time.

회절격자가 집적된 일회용 다중채널 SPR 생체분자 검출 칩 (A Disposable Grating-Integrated Multi-channel SPR Sensor Chip for Detection of Biomolecule)

  • 진영현;조영호
    • 전기학회논문지
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    • 제58권1호
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    • pp.147-154
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    • 2009
  • This paper presents a grating~integrated SPR (Surface Plasmon Resonance) sensor chip for simple and inexpensive biomolecule detection. The grating-integrated SPR sensor chip has two sensing channels having a nano grating for SPR coupling. An external mirror is used for multi channel SPR sensing. The present sensor chip replaces bulky and expensive optical components, such as fiber-optic switches or special shaped prisms, resulting in a simple and inexpensive wavelength modulated multi-channel SPR sensing system. We fabricate a SPR sensor chip integrated with 835 nm-pitch gratings by a micromolding technique to reduce the fabrication cost. In the experimental characterization, the refractive index sensitivity of each sensing channel is measured as $321.8{\pm}8.1nm$/RI and $514.3{\pm}8.lnm$/RI, respectively. 0.5uM of the target biomolecule (streptavidin) was detected by a $1.13{\pm}0.16nm$ shift of the SPR dip in the 10%-biotinylated sample channel, while the SPR dip in the reference channel for environmental perturbation monitoring remained at the same position. From the experimental results, multi-channel biomolecule detection capability of the present grating-integrated SPR sensor chip has been verified. On the basis of the preliminary experiments, we successfully measured the binding reaction rate for the $2\;nM{\sim}200\;nM$ monoclonal-antibiotin, thus verifying biomolecule concentration detectability of the present SPR sensor chip. The binding reaction rates measured from the present SPR sensor chip agredd well with those from a commercialized SPR sensor.

표면 플라즈몬 현미경을 이용한 자기조립 단분자막의 이미징 (Imaging of self-assembled monolayers by surface plasmon microscope)

  • 표현봉;신용범;윤현철;양해식;김윤태
    • 한국광학회지
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    • 제14권1호
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    • pp.97-102
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    • 2003
  • 이차원 표면 플라즈몬의 공명 흡수와 포토 마스크를 이용하여 11-MUA(11-Mercaptoundecanoic acid)와 11-MUOH(11-Mercaptoundecanol) 둥으로 이루어진 자기조립 단분자막(Self-Assembled Monolayer; SAM)의 다채널 영상을 얻었다. 통상의 Photoresist를 이용한 리토그래피 대신에 Thiol bonding의 광산화를 이용하여 패터닝 과정을 줄이고, 백색광 및 대역통과 필터(λ$_{0}$=633nm)를 이용하여 입사광으로써 레이저를 사용할 때 나타나는 간섭무늬를 줄였다. 이로부터 나타나는 이차원 영상의 명암을 정량적으로 보정하면 수 나노미터(nm) 두께의 변화를 측정할 수 있다. 또한 표면 플라즈몬 공명법은 국소화된 근접장 (소산장)을 이용하는 방법으로서, 통상 많이 이용되는 형광법 등에서 나타나는 광탈색(Photobleaching)이나 소광(Quenching) 현상이 없이 시료의 처리가 간단하고, 영상 신호의 시간에 따른 변화가 극히 적으며, 실시간으로 신호의 변화를 측정할 수 있다는 장점이 있다.

A Novel Photonic Crystal Fiber Sensor with Three D-shaped Holes Based on Surface Plasmon Resonance

  • Bing, Pibin;Sui, Jialei;Huang, Shichao;Guo, Xinyue;Li, Zhongyang;Tan, Lian;Yao, Jianquan
    • Current Optics and Photonics
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    • 제3권6호
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    • pp.541-547
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    • 2019
  • A novel photonic crystal fiber (PCF) sensor with three D-shaped holes based on surface plasmon resonance (SPR) is analyzed in this paper. Three D-shaped holes are filled with the analyte, and the gold film is deposited on the side of three planes. The design of D-shaped holes with outward expansion can effectively solve the uniformity problem of metallized nano-coating, it is beneficial to the filling of the analyte and is convenient for real-time measurement of the analyte. Compared with the hexagonal lattice structure, the triangular arrangement of the clad air holes can significantly reduce the transmission loss of light and improve the sensitivity of the sensor. The influences of the air hole diameter, the distance between D-shaped holes and core, and the counterclockwise rotation angle of D-shaped holes on sensing performance are studied. The simulation results show that the wavelength sensitivity of the designed sensor can be as high as 10100 nm/RIU and the resolution can reach 9.9 × 10-6 RIU.

유전체 다중층을 이용한 국소 표면 플라즈몬 공명 센서의 감도 향상에 관한 연구 (Estimation of Sensitivity Enhancements on Localized Surface Plasmon Resonance Sensor Using Dielectric Multilayer)

  • 안희상;강태영;오진우;김규정
    • 한국광학회지
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    • 제28권1호
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    • pp.28-32
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    • 2017
  • 본 논문에서는 LSPR 센서에 적용하기 위한 제한된 높이 100 nm에서 $TiO_2$, $SiO_2$의 다중층을 이용한 LSPR 센서를 디자인을 제안했다. LSPR 센서의 구조는 유전체 층과 나노 구조가 있는 금속층으로 디자인 하였다. 금속층은 금 박막 40 nm와 높이 40 nm, 주기 600 nm, 선폭 300 nm인 나노와이어 구조체를 올려놓은 구조로 디자인하였다. 유전체 층의 높이를 100nm로 제한하고, $TiO_2$, $SiO_2$가 반복되는 구조로 하여 반복층의 개수를 1~4개로 변경하면서 비교 분석하였다. 파장 가변형 SPR을 디자인하기 위해 각도를 75도로 고정하고 파장을 변화시켜 FEM방식으로 계산하였다. 결과로 굴절율이 고정되어 있을 때 다중층의 개수가 증가할수록 공명 파장이 짧아지는 현상을 확인 하였고, 파장의 변화에 더 민감하게 변화하는 것을 측정하였다. 다만, 다중층의 개수가 3개층 이상이 되면 변화하지 않는 것을 확인하였다.

Evaluation of antioxidant property of heat shock protein 90 from duck muscle

  • Zhang, Muhan;Wang, Daoying;Xu, Xinglian;Xu, Weimin
    • Animal Bioscience
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    • 제34권4호
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    • pp.724-733
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    • 2021
  • Objective: The objectives of this study were to investigate the direct antioxidative effect of 90 Kda heat shock protein (Hsp90) obtained from duck muscle. Methods: The interaction of Hsp90 with phospholipids and oxidized phospholipids was studied with surface plasmon resonance (SPR), and their further oxidation in the presence of Hsp90 was evaluated with thiobarbituric acid reactive substances (TBARS) assay. The scavenging effect on the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis (3-ethylbenzthiazoline-6-sulfonic acid (ABTS) was measured, and the electron paramagnetic resonance (EPR) spectroscopy in combination with 5-tert-Butoxycarbonyl-5-methyl-1-pyrroline-N-oxide and 2-phenyl-4,4,5,5,-tetramethylimidazoline-1-oxyl-3-oxide (PTIO) was utilized to determine the abilities of Hsp90 in scavenging hydroxyl and PTIO radicals. Results: SPR showed Hsp90 could bind with both phospholipids and oxidized phospholipids, and prevent their further oxidation by the TBARS assay. The DPPH and ABTS scavenging activity increased with Hsp90 concentration, and could reach 27% and 20% respectively at the protein concentration of 50 μM. The EPR spectra demonstrated Hsp90 could directly scavenge ·OH and PTIO· radicals. Conclusion: This suggests that Hsp90, a natural antioxidant in meat, may play an important role in cellular defense against oxidative stress, and may have potential use in meat products.

A Novel Anticoagulant Protein with High Affinity to Blood Coagulation Factor Va from Tegillarca granosa

  • Jung, Won-Kyo;Jo, Hee-Yeon;Qian, Zhong-Ji;Jeong, Young-Ju;Park, Sae-Gwang;Choi, Il-Whan;Kim, Se-Kwon
    • BMB Reports
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    • 제40권5호
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    • pp.832-838
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    • 2007
  • A novel inhibitory protein against blood coagulation factor Va (FVa) was purified from muscle protein of granulated ark (Tegillarca granosa, order Arcoida, marine bivalvia) by consecutive FPLC method using anion exchange and gel permeation chromatography. In the results of ESI-QTOF tandem mass analysis and database research, it was revealed that the purified T. granosa anticoagulant protein (TGAP) has 7.7 kDa of molecular mass and its partial sequence, HTHLQRAPHPNALGYHGK, has a high identity (64%) with serine/threonine kinase derived from Rhodopirellula baltica (order Planctomycetales, marine bacteria). TGAP could potently prolong thrombin time (TT), corresponding to inhibition of thrombin (FIIa) formation. Specific factor inhibitory assay showed that TGAP inhibits FVa among the major components of prothrombinase complex. In vitro assay for direct-binding affinity using surface plasmon resonance (SPR) spectrometer indicated that TGAP could be directly bound with FVa. In addition, the binding affinity of FVa to FII was decreased by addition of TGAP in dose-dependant manner ($IC_{50}$ value = 77.9 nM). These results illustrated that TGAP might interact with a heavy chain of FVa ($FVa_H$) bound to FII in prothrombin complex. The present study elucidated that non-cytotoxic T. granosa anticoagulant protein (TGAP) bound to FVa can prolong blood coagulation time by inhibiting conversion of FII to FIIa in blood coagulation cascade. In addition, TGAP did not significantly (P < 0.05) show fibrinolytic activity and cytotoxicity on venous endothelial cell line (ECV 304).

Volatile Organic Compound Specific Detection by Electrochemical Signals Using a Cell-Based Sensor

  • Chung, Sang-Gwi;Kim, Jo-Chun;Park, Chong-Ho;Ahn, Woong-Shick;Kim, Yong-Wan;Choi, Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • 제18권1호
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    • pp.145-152
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    • 2008
  • A cell-based in vitro exposure system was developed to determine whether oxidative stress plays a role in the cytotoxic effects of volatile organic compounds (VOCs) such as benzene, toluene, xylene, and chlorobenzene, using human epithelial HeLa cells. Thin films based on cysteine-terminated synthetic oligopeptides were fabricated for immobilization of the HeLa cells on a gold (Au) substrate. In addition, an immobilized cell-based sensor was applied to the electrochemical detection of the VOCs. Layer formation and immobilization of the cells were investigated with surface plasmon resonance (SPR), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). The adhered living cells were exposed to VOCs; this caused a change in the SPR angle and the VOC-specific electrochemical signal. In addition, VOC toxicity was found to correlate with the degree of nitric oxide (NO) generation and EIS. The primary reason for the marked increase in impedance was the change of aqueous electrolyte composition as a result of cell responses. The p53 and NF-${\kappa}B $ downregulation were closely related to the magnitude of growth inhibition associated with increasing concentrations of each VOC. Therefore, the proposed cell immobilization method, using a self-assembly technique and VOC-specific electrochemical signals, can be applied to construct a cell microarray for onsite VOC monitoring.