• Title/Summary/Keyword: SPR biosensor

Search Result 32, Processing Time 0.036 seconds

Nano-scale Probe Fabrication Using Self-assembly Technique and Application to Detection of Escherichia coli O157:H7

  • Oh, Byung-Keun;Lee, Woochang;Lee, Won-Hong;Park, Jeong-Woo
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.8 no.4
    • /
    • pp.227-232
    • /
    • 2003
  • A self-assembled monolayer of protein G was fabricated to develop an immunosensor based on surface plasmon resonance (SPR), thereby improving the performance of the antibodybased biosensor through immobilizing the antibody molecules (lgG). As such, 11-mercaptoundecanoic acid (11-MUA) was adsorbed on a gold (Au) support, while the non-reactive hydrophilic surface was changed through substituting the carboxylic acid group (-COOH) in the 11-MUA molecule using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrocholide (EDAC). The formation of the self-assembled protein G layer on the Au substrate and binding of the antibody and antigen were investigated using SPR spectroscopy, while the surface topographies of the fabricated thin films were analyzed using atomic force microscopy (AFM). A fabricated monoclonal antibody (Mab) layer was applied for detecting E. coli O157:H7. As a result, a linear relationship was achieved between the pathogen concentration and the SPR angle shift, plus the detection limit was enhanced up to 10$^2$ CFU/mL.

Estimation of Sensitivity Enhancements of Material-Dependent Localized Surface Plasmon Resonance Sensor Using Nanowire Patterns (금속물질에 따른 나노구조를 이용한 국소 표면 플라즈몬 공명 센서 특성 분석)

  • Ahn, Heesang;Ahn, Dong-Gyu;Song, Yung Min;Kim, Kyujung
    • Journal of the Korean Society for Precision Engineering
    • /
    • v.33 no.5
    • /
    • pp.363-369
    • /
    • 2016
  • We explored localized plasmonic field enhancements using nanowire patterns to improve the sensitivity of a surface plasmon resonance (SPR) sensor. Two different materials, gold and silver, were considered for sample materials. Gold and silver nanowire patterns were fabricated by electron beam lithography for experimental measurements. The wavelength SPR sensor was also designed for these experiments. The material-dependent field enhancements on nanowire patterns were first calculated based on Maxwell's equations. Resonance wavelength shifts were indicated as changes in the refractive index from 1.33 to 1.36. The SPR sensor with silver nanowire patterns showed a much larger resonance wavelength shift than the sensor with gold nanowire patterns, in good agreement with simulation results. These results suggest that silver nanowire patterns are more efficient than gold nanowire patterns, and could be used for sensitivity enhancements in situations where biocompatibility is not a consideration.

Pre-processing Algorithm for Analysis from Microarray Image Sequence by Surface Plasmon Resonance Biosensor (SPR 센서로 촬영된 마이크로어레이 영상 분석을 위한 전처리 알고리즘)

  • 황지운;김재학;한준희
    • Proceedings of the Korean Information Science Society Conference
    • /
    • 2004.04b
    • /
    • pp.250-252
    • /
    • 2004
  • 나노 기술의 개발과 마이크로어레이(microarray)의 등장으로 생물학자동을 한번에 보다 라르고 신속하게, 대량의 실험을 처리할 수 있게 되었다. 이와 발 맞추어 마이크로어레이를 이용한 다양한 실험 방법들이 개발되었다. 형광 염료(fluorescence dye)를 이 용한 관찰 방법 이 널리 이용되고 있으나, 관찰되는 형광 염료의 밝기가 실험 환경(pH, 온도)에 매우 민감하게 반응하며, 단백질을 포함한 많은 분자 물질들이 형광을 내지 않기 때문에 마이크로어레이를 이용한 분석 대상 물질들의 개수가 제한을 받는다. 본 논문에서는 직접적인 형광 염료의 사용 없이, SPR(Surface Plasmon Resonance)을 이용한 마이크로어레이 분석 시스템에서 스팟(spot)의 밝기(intensity)를 측정하기 위한 효율적인 전처리 과정을 제안하고자 한다. 전처리 과정은 크게 프로젝티브 왜곡 효과 제거, 스팟의 위치 추적, 스팟의 영역 추출, 정규화 된 스팟의 밝기 측정으로 나누어진다. 특히, 이러한 과정을 거쳐서 측정된 밝기는 반응 유무의 관찰뿐만이 아니라, 실험 물질의 양적인 측정에도 이용되기 때문에 정확한 스팟의 밝기 측정에 중점을 두고자 한다.

  • PDF

Detection of deoxynivalenol using a MOSFET-based biosensor (MOSFET형 바이오 센서를 이용한 디옥시 니발레놀의 검출)

  • Lim, Byoung-Hyun;Kwon, In-Su;Lee, Hee-Ho;Choi, Young-Sam;Shin, Jang-Kyoo;Choi, Sung-Wook;Chun, Hyang-Sook
    • Journal of Sensor Science and Technology
    • /
    • v.19 no.4
    • /
    • pp.306-312
    • /
    • 2010
  • We have detected deoxynivalenol(DON) using a metal-oxide-semiconductor field-effect-transistor(MOSFET)-based biosensor. The MOSFET-based biosensor is fabricated by a standard complementary metal-oxide-semiconductor(CMOS) process, and the biosensor's electrical characteristics were investigated. The output of the sensor was stabilized by employing a reference electrode that applies a fixed bias to the gate. Au which has a chemical affinity for thiol was used as the gate metal to immobilize a self-assembled monolayer(SAM) made of 16-mercaptohexadecanoic acid(MHDA). The SAM was used to immobilize anti-deoxynivalenol antibody. The carboxyl group of the SAM was bound to the anti- deoxynivalenol antibody. Anti-deoxynivalenol antibody and deoxynivalenol were bound by an antigen-antibody reaction. In this study, it is confirmed that the MOSFET-based biosensor can detect deoxynivalenol at concentrations as low as 0.1 ${\mu}g$/ml. The measurements were performed in phosphate buffered saline(PBS; pH 7.4) solution. To verify the interaction among the SAM, antibody, and antigen, surface plasmon resonance(SPR) measurements were performed.

Strategies in Protein Immobilization on a Gold Surface

  • Park, Jeho;Kim, Moonil
    • Applied Science and Convergence Technology
    • /
    • v.24 no.1
    • /
    • pp.1-8
    • /
    • 2015
  • Protein immobilization on a gold surface plays an important role in the usefulness of biosensors that utilize gold-coated surfaces such as surface plasmon resonance (SPR), quartz crystal microbalance (QCM), etc. For developing high performance biosensors, it is necessarily required that immobilized proteins must remain biologically active. Loss of protein activity and maintenance of its stability on transducer surfaces is directly associated with the choice of immobilization methods, affecting protein-protein interactions. During the past decade, a variety of strategies have been extensively developed for the effective immobilization of proteins in terms of the orientation, density, and stability of immobilized proteins on analytical devices operating on different principles. In this review, recent advances and novel strategies in protein immobilization technologies developed for biosensors are briefly discussed, thereby providing an useful information for the selection of appropriate immobilization approach.

Label-free Detection of Biomolecular Specific Interaction by Optical Biosensors (광 바이오센서를 이용한 비표지 생계물질들의 특이 상호작용력의 측정)

  • 김의락;최정우
    • KSBB Journal
    • /
    • v.17 no.1
    • /
    • pp.1-13
    • /
    • 2002
  • Label-free optical methods for the monitoring of interactions between biological molecules have become increasingly popular within the last decade. A rising number of publications have demonstrated the benefits of direct biomolecular interaction analysis(BIA) for biology and biochemistry, such as antigen-antibody Interactions, receptor-ligand interactions, protein-DNA, DNA- intercalator, and DNA-DNA interactions. This article gives an overview of the historical development, principle and application of label-free optical biosensor to examine the functional characteristics of biospecific interaction, such as kinetics, affinity, and binding position of biomolecular between an immobilized species at the transducer surface and its dissolved binding partner.

Receptor Binding Affinities of Synthetic Cannabinoids Determined by Non-Isotopic Receptor Binding Assay

  • Cha, Hye Jin;Song, Yun Jeong;Lee, Da Eun;Kim, Young-Hoon;Shin, Jisoon;Jang, Choon-Gon;Suh, Soo Kyung;Kim, Sung Jin;Yun, Jaesuk
    • Toxicological Research
    • /
    • v.35 no.1
    • /
    • pp.37-44
    • /
    • 2019
  • A major predictor of the efficacy of natural or synthetic cannabinoids is their binding affinity to the cannabinoid type I receptor ($CB_1$) in the central nervous system, as the main psychological effects of cannabinoids are achieved via binding to this receptor. Conventionally, receptor binding assays have been performed using isotopes, which are inconvenient owing to the effects of radioactivity. In the present study, the binding affinities of five cannabinoids for purified $CB_1$ were measured using a surface plasmon resonance (SPR) technique as a putative non-isotopic receptor binding assay. Results were compared with those of a radio-isotope-labeled receptor binding assay. The representative natural cannabinoid ${\Delta}^9$-tetrahydrocannabinol and four synthetic cannabinoids, JWH-015, JWH-210, RCS-4, and JWH-250, were assessed using both the SPR biosensor assay and the conventional isotopic receptor binding assay. The binding affinities of the test substances to $CB_1$ were determined to be (from highest to lowest) $9.52{\times}10^{-3}M$ (JWH-210), $6.54{\times}10^{-12}M$ (JWH-250), $1.56{\times}10^{-11}M$ (${\Delta}^9$-tetrahydrocannabinol), $2.75{\times}10^{-11}M$ (RCS-4), and $6.80{\times}10^{-11}M$ (JWH-015) using the non-isotopic method. Using the conventional isotopic receptor binding assay, the same order of affinities was observed. In conclusion, our results support the use of kinetic analysis via SPR in place of the isotopic receptor binding assay. To replace the receptor binding affinity assay with SPR techniques in routine assays, further studies for method validation will be needed in the future.

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.

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.