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

검색결과 126건 처리시간 0.025초

전사인자 저해제 통한 미백제 탐색용 단백질 칩 제작 (Manufacturing Protein-DNA Chip for Depigmenting Agent Screening)

  • 한정선;곽은영;이향복;신정현;백승학;정봉현;김은기
    • 대한화장품학회지
    • /
    • 제30권4호
    • /
    • pp.479-483
    • /
    • 2004
  • MITF는 미백관련 유전자의 대표적인 조절 인자 단백질로서 미백관련 유전자의 E-box와의 결합정도를 단백질 칩을 이용하여 측정하였다. 융합 단백질 형태의 MITF를 유리 칩에 고정시켰고 E-box를 포함하는 DNA oligomer가 결합하는 것을 확인하였다. 형광법, SPR (surface plasmon resonance), SPRi (surface plasmon resonance imaging)방법 중 형광법이 가장 효과적이었으며, DNA 저해제를 사용시 결합이 감소하는 것을 확인하였다. 이 결과 MITF를 이용한 미백원료의 고속스크리닝(HTS)의 가능성을 보여주었다.

Characterization of binding specificity using GST-conjugated mutant huntingtin epitopes in surface plasmon resonance (SPR)

  • Cho, Hang-Hee;Kim, Tae Hoon;Kim, Hong-Duck;Cho, Jae-Hyeon
    • 한국동물위생학회지
    • /
    • 제44권4호
    • /
    • pp.185-194
    • /
    • 2021
  • Polyglutamine extension in the coding sequence of mutant huntingtin causes neuronal degeneration associated with the formation of insoluble polyglutamine aggregates in Huntington's disease (HD). Mutant huntingtin can form aggregates within the nucleus and processes of neurons possibly due to misfolding of the proteins. To better understand the mechanism by which an elongated polyglutamine causes aggregates, we have developed an in vitro binding assay system of polyglutamine tract from truncated huntingtin. We made GST-HD exon1 fusion proteins which have expanded polyglutamine epitopes (e.g., 17, 23, 32, 46, 60, 78, 81, and 94 CAG repeats). In the present emergence of new study adjusted nanotechnology on protein chip such as surface plasmon resonance strategy which used to determine the substance which protein binds in drug discovery platform is worth to understand better neurodegenerative diseases (i.e., Alzheimer disease, Parkinson disease and Huntington disease) and its pathogenesis along with development of therapeutic measures. Hence, we used strengths of surface plasmon resonance (SPR) technology which is enabled to examine binding specificity and explore targeted molecular epitope using its electron charged wave pattern in HD pathogenesis utilize conjugated mutant epitope of HD protein and its interaction whether wild type GST-HD interacts with mutant GST-HD with maximum binding affinity at pH 6.85. We found that the maximum binding affinity of GST-HD17 with GST-HD81 was higher than the binding affinities of GST-HD17 with other mutant GST-HD constructs. Furthermore, our finding illustrated that the mutant form of GST-HD60 showed a stronger binding to GST-HD23 or GST-HD17 than GST-HD60 or GST-HD81. These results indicate that the binding affinity of mutant huntingtin does not correlate with the length of polyglutamine. It suggests that the aggregation of an expanded polyglutamine might have easily occurred in the presence of wild type form of huntingtin.

Characterization of biotin-avidin recognition system constructed on the solid substrate

  • Lim, Jung-Hyurk
    • 분석과학
    • /
    • 제18권6호
    • /
    • pp.460-468
    • /
    • 2005
  • The biotin-avidin complex, as a model recognition system, has been constructed through N-hydroxysuccinimide(NHS) reaction on a variety of substrates such as a smooth Au film, electrochemically roughened Au electrode and chemically modified mica. Stepwise self-assembled monolayers (SAMs) of biotin-avidin system were characterized by surface-enhanced resonance Raman scattering (SERRS) spectroscopy, atomic force microscopy (AFM) and surface plasmon resonance (SPR). A strong SERRS signal of rhodamine tags labeled in avidin from the SAMs on a roughened gold electrode indicated the successful complex formation of stepwise biotin-avidin recognition system. AFM images showed the circular shaped avidin aggregates (hexamer) with ca. $60{\AA}$ thick on the substrate, corresponding to one layer of avidin. The surface coverage and concentration of avidin molecules were estimated to be 90% and $7.5{\times}10^{-12}mol/cm^2$, respectively. SPR technique allowed one to monitor the surface reaction of the specific recognition with high sensitivity and precision.

단백질 칩을 이용한 클라미디아 폐렴의 진단 (Development of Protein Chip for Diagnosis of Chlamydophia Pneumoniae)

  • 김우진;이희영;이승준;정세희;육종설;하권수;정기석
    • Tuberculosis and Respiratory Diseases
    • /
    • 제60권4호
    • /
    • pp.412-418
    • /
    • 2006
  • 연구배경 : 클라미디아 감염의 진단은 혈청검사로 이루어진다. 현재 표준 방법은 MIF(microimmunofluorescence)이나 이 방법은 주관적이고 시간이 많이 걸리는 단점이 있다. 최근을 SPR(surface plasmon resonance) 센서를 이용한 단백질 칩이 감염의 새로운 진단 방법으로 제시되고 있다. 클라미디아 감염의 진단을 위한 단백질 칩 개발을 위하여 금 칩 표면에 세균을 고정하고 클라미디아 균에 대한 항체와 표면 위 세균과의 반응을 SPR 센서를 이용하여 측정하고자 하였다. 방법 : 표면 항원으로 배양한 Chlamydophila pneumoniae LKK1의 EB를 정제하였다. 양전하를 띤 PDDA (polydiallyldimethylammonium chloride)를 이용하여 전하를 이용한 단백질 칩을 제작하였다. 클라미디아 균을 고정시킨 후에 atomic force microscopy를 이용하여 표면을 관찰하였다. 클라미디아 균에 대한 항체를 투여하고 나서 자체 제작한 SPR 센서를 이용하여 항원 항체 반응을 SPR 파장 변화로 측정하였다. 결과 : 양전하를 띤 PDDA 표면위에서 클라미디아 균이 고정되었음을 확인 하였다. 그리고, 항체를 투여한 후에 SPR 파장의 증가를 확인하였다. 파장 변화는 항원의 농도와 관련이 있었다. 결론 : 전하를 이용하여 클라미디아 폐렴균의 EB를 단백질 칩에 고정하였고, 단백질 칩 위에서의 항원 항체 반응을 확인하였다. 비정형 폐렴의 진단에 SPR 센서가 기여할 수 있을 것으로 사료되나, 실제 임상 시료에의 적용을 위해서는 좀더 연구가 필요할 것으로 사료된다.

Passivation of organic light emitting diodes with $Al_2O_3/Ag/Al_2O_3$ multilayer thin films grown by twin target sputtering system

  • Jeong, Jin-A;Kim, Han-Ki
    • 한국정보디스플레이학회:학술대회논문집
    • /
    • 한국정보디스플레이학회 2008년도 International Meeting on Information Display
    • /
    • pp.420-423
    • /
    • 2008
  • The characteristics of $Al_2O_3/Ag/Al_2O_3$ multilayer passivaton prepared by twin target sputtering (TTS) system for organic light emitting diodes. The $Al_2O_3/Ag/Al_2O_3$ multilayer thin film passivation on a PET substrate had a high transmittance of 86.44 % and low water vapor transmission rate (WVTR) of $0.011\;g/m^2$-day due to the surface plasmon resonance (SPR) effect of Ag interlayer and effective multilayer structure for preventing the intrusion of water vapor. Using synchrotron x-ray scattering and field emission scanning electron microscope (FESEM) examinations, we investigated the growth behavior of Ag layer on the $Al_2O_3$ layer to explain the SPR effect of the Ag layer. This indicates that an $Al_2O_3/Ag/Al_2O_3$ multilayer passivation is a promising thin film passivation scheme for organic based flexible optoelectronics.

  • PDF

생체분자와 필드의 동시국소화를 통한 플라스몬 센서의 감도향상 연구 (Enhanced Detection Sensitivity of Surface Plasmon Resonance Biosensing Based on Colocalized Target Molecules and Evanescent Fields)

  • 이원주;오영진;김동현
    • 한국광학회지
    • /
    • 제22권4호
    • /
    • pp.198-203
    • /
    • 2011
  • 본 논문에서는 국소화된 플라스몬 필드 내에 분자를 동시국소화 시키는 방법으로 플라스몬 센서의 감도를 향상시키기 위한 이론적 연구를 수행하였다. 플라스몬 필드의 국소화는 나노격자를 통하여 이루어 졌으며, 측정하고자 하는 분자 반응은 유전체 박막의 기울임 증착을 통하여 국소화되는 것으로 가정하였다. 근접장 기반의 필드와 분자 분포 간의 중복적분 값을 통하여 중복도가 플라스몬 센서의 감도와 밀접한 관련이 있는 것을 확인하였으며, 계산된 플라스몬 센서 특성에 근거하여 동시국소화된 플라스몬 센서의 경우, 동시국소화되지 않은 경우에 비하여, 10배 정도의 감도개선 효과가 있는 것을 확인하였다.

실리카 코어 금 위성입자의 합성 및 표면 증강 라만 산란을 기반으로 한 센서로의 응용 (Synthesis of Silica-Core Gold-Satellite Nanoparticles and Their Surface-enhanced Raman Scattering Based Sensing Application)

  • 최현지;김영국;윤석영;백연경
    • 한국분말재료학회지
    • /
    • 제21권6호
    • /
    • pp.441-446
    • /
    • 2014
  • In this study, we synthesize silica-core gold-satellite nanoparticles (SGNPs) for the surface-enhanced Raman scattering (SERS) based sensing applications. They consist of gold satellite nanoparticles (AuNPs) fixed on the silica core nanoparticles, which sizes of AuNPs can be tunned by varying the amount of reactants (growth solution and reducing agent). Their surface plasmon resonance (SPR) properties were characterized by using UV-vis spectroscopy, showing that the growth of AuNPs on silica cores leads to the light absorption in the longer wavelength region. Furthermore, the size increase of AuNPs exhibited the dramatic change in SERS activity due to the formation of hot spots. The optimized SGNPs showing enhancement factor ${\sim}3.8{\times}10^6$ exhibited a detection limit of rhodamine 6G (R6G) as low as $10^{-8}M$. These findings suggest the importance of size control of SGNPs and their SPR properties to develop highly efficient SERS sensors.

Surface plasmon resonance 바이오센서를 이용한 재조합 B형 간염 표면항원의 정량분석 (Quantitative Assay of Recombinant Hepatitis B Surface Antigen by Using Surface Plasmon Resonance Biosensor)

  • 이은규;안상점;유창훈;류강;전준영;이현익;최성철;이영식
    • KSBB Journal
    • /
    • 제17권1호
    • /
    • pp.20-25
    • /
    • 2002
  • We performed a basic experiment for rapid, on-line, real-time measurement of HBsAg by using a surface plasmon resonance biosensor to quantify the recognition and interaction of biomolecules. We immobilized the anti-HBsAg polyclonal antibody to the dextran layer on a CM5 chip surface which was pre-activated by N-hydroxysuccinimide for amine coupling. The binding of the HBsAg to the immobilized antibody was measured by the mass increase detected by the change in the SPR signal. The binding characteristics between HBsAg and its antibody followed typical monolayer adsorption isotherm. When the entire immobilized antibody was interacted, there was no additional, non-specific binding observed, which suggested the biointeraction was very specific as expected and independent of the ligand density. No significant steric hindrance was observed at 17.6 nm/$mm^2$ immobilization density. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the chip surface was linear up to ca. $40\mu\textrm{g}$/mL, which is much wider than that of the ELISA method. It appeared the antigen-antibody binding was increased as the immobilized ligand density increased, but verification is warranted. This study showed the potential of this biosensor-based method as a rapid, simple, multi-sample, on-line assay. Once properly validated, it can serve as a more powerful method for HBsAg quantification replacing the current ELISA method.

표면플라즈몬공명과 효소면역분석법을 이용한 살충제 카보후란 잔류물 검출 (Sensing of the Insecticide Carbofuran Residues by Surface Plasmon Resonance and Immunoassay)

  • 양길모;조남홍
    • Journal of Biosystems Engineering
    • /
    • 제30권6호통권113호
    • /
    • pp.333-339
    • /
    • 2005
  • The pesticide is raising public interest in the world, because it causes damage to an environmental pollution and the human health remaining agricultural products and an ecosystem, in spite of the advantages. Particularly, each country restricts the residual pesticide and induces observance about the safety and usage standard so that they can control the amount of pesticide used and defend the safety of agricultural products. The habitual practice for the analysis of the residual pesticide depends on GC (gas chromatography), HPLC (high performance liquid chromatography) and GC/MS (gas chromatography/mass spectroscopy), which triturate the fixed quantity of samples, abstract and purify as a suitable organic solvent. These methods have the highly efficient in aspects of sensitivity and accuracy. On the other hand, they need the high cost, time consuming, much effort, expensive equipment and the skillful management. Carbofuran is highly toxic by inhalation and ingestion and moderately toxic by dermal absorption. As with other carbamate compounds, it is metabolized in the liver and eventually excreted in the urine. The half-life of carbofuran on crops is about 4 days when applied to roots, and longer than 4 days if applied to the leaves. This research was conducted to develop immunoassay for detecting carbofuran residue quickly on the basis of surface plasmon resonance and to evaluate the measurement sensitivity. Gold chip used was CM5 spreaded dextran on the surface. An applied antibody to Immunoassay was GST (glutathione-s-transferase). The association and the dissociation time were 176 second and 215 second between GST and carbofuran. The total analysis time using surface plasmon resonance was 13 minutes including regeneration time, on the other hand HPLC and GC/MS was 2 hours usually. The minimum detection limit of a permissible amount for carbofuran in the country is 0.1 ppm. The immunoassay method using surface plasmon resonance was 0.002 ppm.

Aggregation of Laser-Generated Gold Nanoparticles Mediated by Formalin

  • Alauddin, Md.;Kim, Kuk Ki;Roy, Madhusudan;Song, Jae Kyu;Kim, Myung Soo;Park, Seung Min
    • Bulletin of the Korean Chemical Society
    • /
    • 제34권1호
    • /
    • pp.188-196
    • /
    • 2013
  • We have investigated the effects of formalin on the assembly of colloidal gold nanoparticles (AuNPs) prepared by laser ablation of a solid gold target in deionized water. Upon addition of formalin, the surface plasmon resonance (SPR) band at 519 nm for pure AuNPs decreases and shifts to red while a new broad SPR band appears at ~700 nm. The red-shift is prominent with increase in the incubation time. The average size of the initial AuNPs is around 12 nm but it increases to 23 nm after addition of formalin. It turns out that formalin acts as a cationic surfactant for AuNPs with negative surface charge in the colloidal solutions. Furthermore, through analysis of the Raman spectrum of formalin and the density functional theory calculations, we confirm that methanediol is the main species in formalin which is in charge of the aggregation of AuNPs.