• Title/Summary/Keyword: Surface plasmon resonance(SPR)

Search Result 126, Processing Time 0.025 seconds

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

  • Han Jung-Sun;Kwak Eun-Young;Lee Hyang-Bok;Shin Jlung-Hyun;Baek Seung-Hak;Chung Bong-Hyun;Kim Eun-Ki
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.30 no.4 s.48
    • /
    • pp.479-483
    • /
    • 2004
  • An attempt was made to develop a proteinchip for screening of MITF (microphthalmia transcription factor) inhibitor. Binding of MITF to E-box causes transcription of several pigmenting genes including tyrosinase gene. We investigated binding of MITF and its DNA binding site (E-box) using a protein-DNA chip with various detection methods including flurorescence (Cyt3). SPR (surface plasmon resonance) and SPRi (surface plasmon resonance imaging). A fusion protein (MITF-Maltose Binding Protein) was attached on the glass plate by chemical modification. An inhibitory synthetic DNA oligomer, artificially designed based on the E-box sequence, inhibited the binding of MITF and E-box. These results showed the potentials of flurorescence-based MITF protein chip as a microarray for high throughput screening (HTS) system of depigmenting agents.

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
    • Korean Journal of Veterinary Service
    • /
    • v.44 no.4
    • /
    • 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
    • Analytical Science and Technology
    • /
    • v.18 no.6
    • /
    • 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.

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

  • Kim, Woo Jin;Lee, Hui Young;Lee, Seung-Joon;Jung, Se-Hui;Yuk, Jong Seol;Ha, Kwon-Soo;Jung, Ki-Suck
    • Tuberculosis and Respiratory Diseases
    • /
    • v.60 no.4
    • /
    • pp.412-418
    • /
    • 2006
  • Background; The diagnosis of chlamydial infection is based on serology. The current gold standard of diagnosis is MIF(microimmunofluorescence), but this modality is subjective and time-consuming. Protein microarray with using a SPR(surface plasmon resonance) sensor has recently been suggested as a method for detecting infection. For developing a protein chip to diagnose chlamydial infection, EBs(elementary bodies) were immobilized on a gold chip and the interaction between an antibody for Chlamydophila pneumoniae and the EBs(elementary bodies) immobilized on the surface of the gold chip was measured by using an SPR sensor. Methods; For the surface antigen, the EBs of Chlamydophila pneumoniae LKK1 were purified. Charged arrays were prepared by using PDDA(polydiallyldimethylammonium chloride) which has a positive charge. After immobilization of the chlamydial EBs on the PDDA surface, the investigation of the surface was done with using atomic force microscopy. After the antibody for C. pneumoniae was applied on chip, we monitored the SPR wavelength-shift to detect any antigen-antibody interaction with using a self-assembled SPR sensor. Results; The chlamydial EBs on the positively charged PDDA were visible on the surface with using atomic force microscopy. The SPR wavelength increased after interaction of antibody for C. pneumoniae with the EBs immobilized on charged gold surface. The wavelength-shift was correlated with the concentration of antigens. Conclusion; The surface immobilization of EBs on the gold surface with the charged arrays was identified and the antigen-antibody interaction on the gold chip was detected via the SPR sensor. Further investigations are needed to apply this technique to the clinical field.

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
    • /
    • 2008.10a
    • /
    • 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

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

  • Lee, Won-Ju;Oh, Young-Jin;Kim, Dong-Hyun
    • Korean Journal of Optics and Photonics
    • /
    • v.22 no.4
    • /
    • pp.198-203
    • /
    • 2011
  • We have conducted a theoretical study to improve the detection limit of a surface plasmon resonance (SPR) sensor by co-localizing plasmonic fields and target molecules of interest. The fields were localized by nanograting antennas, while target molecules that participate in a molecular interaction were assumed to be co-localized by angled evaporation of a dielectric mask layer on the nanograting antennas. We have performed the evaluation using an overlap integral between distributions of plasmon fields and molecules and confirmed the correlation of the overlap with the sensitivity of an SPR sensor. Based on the calculated sensor characteristics, it was found that the sensitivity, if the fields and molecules are co-localized, can be as much as ten times that of non-colocalized structure.

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

  • Choi, Hyun Ji;Kim, Young-Kuk;Yoon, Seok-Young;Baek, Youn-Kyoung
    • Journal of Powder Materials
    • /
    • v.21 no.6
    • /
    • 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.

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

  • Lee, E. K.;Ahn, S. J.;Yoo, C. H.;Ryu, K.;Jeon, J. Y.;Lee, H. I.;Choi, S. C.;Lee, Y. S.
    • KSBB Journal
    • /
    • v.17 no.1
    • /
    • 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.

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

  • Yang G. M.;Cho N. H.
    • Journal of Biosystems Engineering
    • /
    • v.30 no.6 s.113
    • /
    • 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
    • /
    • v.34 no.1
    • /
    • 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.