• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.4
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• pp.309-314
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• 2005

We performed a basic experiment for the rapid, on-line, real-time measurement of hepatitis B surface antigen using a surface plasmon resonance biosensor. We immobilized anti­HBsAg (hepatitis B surface antigen) polyclonal antibody, as a ligand, to the dextran layer on a CM5 chip surface that had previously been activated by N-hydroxysuccinimide. A sample solution containing HBsAg was fed through a microfluidic channel, and the reflecting angle change due to the mass increase from the binding was detected. The binding characteristics between HBsAg and its polyclonal antibody followed the typical monolayer adsorption isotherm. When the entire immobilized antibody had interacted, no additional, non-specific binding occurred, suggesting the immunoreaction was very specific. The bound antigen per unit mass of the antibody was independent of the immobilized ligand density. No significant steric hindrance was observed at an immobilization density of approximately $17.6 ng/mm^2$. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the ligand was linear up to ca. $40{\mu}g$/mL. This linearity was much higher than that of the ELISA method. It appeared the anti­gen-antibody binding increased as the immobilized ligand density increased. In summary, this study showed the potential of this SPR biosensor-based method as a rapid, simple and multi­sample on-line assay. Once properly validated, it may serve as a more efficient method for HBsAg quantification for replacing the ELISA.

• Journal of Microbiology and Biotechnology
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• v.18 no.1
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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.

• Proceedings of the Korean Vacuum Society Conference
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• 2011.02a
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• pp.426-426
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• 2011

최근 들어 금속물질을 나노미터 단위로 구성할 수 있는 기술이 진보하면서, 금속 나노입자에 의해 발생되는 표면 플라즈몬에 대해서도 다양한 분야의 관심이 집중되고 있다. 유전체 물질을 기지상으로 하는 금속:유전체 나노복합체에서 금속 나노입자는 자유전자들의 집단 진동인 국소표면 플라즈몬 공진(Localized Surface Plasmon Resonance, LSPR)현상에 의해 국부전기장을 증대 시키고, 가시광 및 적외선 영역에서 특성 광흡수 거동을 보인다. 이와 같은 광학적 특성은 금속 나노입자들의 크기, 형태, 그리고 나노입자들의 주변을 구성하는 기지상 물질의 종류에 의해 조절된다. 금속:유전체 나노복합체에 나타나는 이러한 특성은 단순장식코팅 뿐만 아니라 광의 효율적 운용과 광을 매개로 한 기능발현을 필요로 하는 디스플레이, 광학 스위칭 소재 및 태양전지의 효율 향상을 위한 광흡수층 등 매우 다양한 응용이 가능하다. 본 연구에서는 다양한 굴절률을 갖는 재료들 중, 저굴절률을 갖는 SiO2와 고굴절률을 갖는 ZnS-SiO2를 기지상 재료로 선택하여 교번증착 스퍼터링법으로 Ag와 Au입자를 형성시켰다. Ag를 금속나노입자로 갖고, SiO2와 ZnS-SiO2를 기지상으로 하는 금속:유전체 나노복합체에서는 금속나노입자 형성에 따른 뚜렷한 표면 플라즈몬 공진 광흡수 피크가 관찰된 반면 Au나노입자는 기지상에 따라 각기 다른 광흡수 특성을 나타냈는데, SiO2기지상에서 명확한 광흡수 피크를 형성했던 경우와는 달리 ZnS-SiO2기지상에서는 특정파장에서의 흡수피크로 규정되기 어려운 넓은 파장범위에 걸친 완만한 광흡수 피크를 나타냈다. TEM 분석을 통해, ZnS-SiO2 기지상 내의 Au입자는 각각 독립되어 있는 Island형태가 아닌 유전체 기지상과 대칭적으로 혼합된 네트워크 형태의 Bruggeman 기하구조를 구성하고 있음을 확인하였고, 이는 Au입자가 형성되고 성장할 때 Au와 S의 높은 결합에너지로 인해 상당한 젖음 특성을 갖고 성장하였기 때문으로 판단됐다. 따라서 나노복합체를 구성하는 물질간의 광학적 특성뿐만 아니라 기지상 내에서의 금속입자의 성장거동에 대한 연구가 수반되었을 때, 금속:유전체 나노복합체의 표면 플라즈몬 공진 광흡수 특성을 보다 정확하게 제어할 수 있다.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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• 2008.06a
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• pp.76-76
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• 2008

The resonance properties due to the surface plasmon(SP) excitation of metal nanoparticles make the nanocomposite films promising for various applications such as optical switching devices. In spite of the well-known ultra-sensitive operation of optical switches based on a guided wave, the application of nanocomposite film(NC) has inherent limitation originating from the excessive optical loss related with the surface plasmon resonance(SPR). In this study, we addressed this problem and present the experimental and theoretical analysis on the pump-probe optical switching in prism-coupled Au(1 vol.%):$SiO_2$ nanocomposite waveguide film. The guided mode was successfully generated using a near infrared probe beam of 1550 nm and modulated with an external pump beam of 532 nm close to the SPR wavelength. We extend our approach to ultra-fast operation using a pulsed laser with 5 ns pulse width. To improve the switching speed through the reduction in thermal loading effect accompanied by the resonant absorption of pump beam light, we adopted a metallic film as a coupling layer instead of low-index dielectric layer between the high-index SF10 prism and NC slab waveguide. We observed great enhancement in switching speed for the case of using metallic coupling layer, and founded a distinct difference in origin of optical nonlinearities induced during switching operation using cw and ns laser.

• Journal of the Korean Institute of Electrical and Electronic Material Engineers
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• v.28 no.4
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• pp.244-248
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• 2015

We proposed and demonstrated the double layered metallic nano-hole structure using polystyrene beads process to enhance the sensitivity of surface plasmon resonance (SPR). The double layered SPR structures are calculated using the finite-difference time-domain (FDTD) method for the width, thickness, and period of the metallic nano-hole structures. The thickness of the metal film and the metallic nano-hole is 30 and 20 nm in the 214 nm wide nano-hole size, respectively. The double layered SPR structures are fabricated with monolayer polystyrene beads of 420 nm wide. The sensitivities of the conventional SPR sensor and the double layered SPR sensor are obtained to 42.2 and 52.1 degree/RIU, respectively.

• Journal of Life Science
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• v.13 no.4
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• pp.463-472
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• 2003

Interactions between factor Va (HFVa) and membrane phosphatidylserine (PS) regulate the activity of the prothrombinase complex. I have previously shown that two solvent exposed hydrophobic residues located in the C2-domain, Trp2063 and Trp2064, are required for binding to immobilized PS and for expression of procoagulant activity on membranes containing 5% PS. In order to fully define the functional importance of these two residues I have expressed and isolated recombinant factor Va (rHFVa) W2063A/W2064A double mutant. In contrast to the native protein the two glycoforms resulting from alternative glycosylation of Asn2181 eluted as a single peak with rHFVa1 W2063A/W2064A eluting on the leading edge and rHFVa2 W2063A/W2064A eluting on the trailing edge. The double mutant rHFVa2 W2063A/W2064A expressed little or no procoagulant activity on membranes containing 1-10% mol % PS. In contrast, the procoagulant activity of this mutant was slightly greater than the native protein on membranes containing>18 mol % PS. The binding of rHFVa2 W2063A/W2064A to immobilized phospholipid vesicles was markedly reduced compared to the native protein in a surface plasmon resonance binding assay. I conclude that Trp2063 and Trp2064 are required for high affinity binding of factor Va to PS membranes and that this interaction is necessary for assembly of the prothrombinase complex on membranes containing physiological concentrations of PS.

• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.510-518
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• 2021

Bacteriophages are considered excellent sensing elements for platforms detecting bacteria. However, their lytic cycle has restricted their efficacy. Here, we used the minor coat protein pIII domain (N1N2) of phage CTXφ to construct a novel surface plasmon resonance (SPR) biosensor that could detect Vibrio cholerae. N1N2 harboring the domains required for phage adsorption and entry was obtained from Escherichia coli using recombinant protein expression and purification. SDS-PAGE revealed an approximate size of 30 kDa for N1N2. Dot blot and transmission electron microscopy analyses revealed that the protein bound to the host V. cholerae but not to non-host E. coli K-12 cells. Next, we used amine-coupling to develop a novel recombinant N1N2 (rN1N2)-functionalized SPR biosensor by immobilizing rN1N2 proteins on gold substrates and using SPR to monitor the binding kinetics of the proteins with target bacteria. We observed rapid detection of V. cholerae in the range of approximately 10³ to 10⁹ CFU/ml but not of E. coli at any tested concentration, thereby confirming that the biosensor exhibited differential recognition and binding. The results indicate that the novel biosensor can rapidly monitor a target pathogenic microorganism in the environment and is very useful for monitoring food safety and facilitating early disease prevention.

• Journal of the Korean Institute of Electrical and Electronic Material Engineers
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• v.35 no.4
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• pp.392-397
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• 2022

We present the structural and optical properties of Au@TiO₂ core-shell microsphere structure prepared by a hydrothermal synthesis method. As a way to improve the efficiency of organic solar cells, the Au@TiO₂ core-shell microsphere was synthesized to use the local surface plasmon resonance (LSPR) phenomenon. The synthesized results were confirmed to have the Au@TiO₂ core-shell structure using a high-resolution transmission electron microscopy. An absorption was observed to occur at 527 nm belonging to the visible light region using a visible light spectroscopy, which supports the LSPR phenomenon. We suggest that the Au@TiO₂ core-shell microsphere is highly likely to be applied to organic solar cells including dye-sensitized solar cells. In addition, we expect it to be widely used not only in the energy but also in the bio as well as in the environmental fields.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.02a
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• pp.105-106
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• 2013

Surface plasmon resonance (SPR) is classified into the propagating surface plasmon (PSP) excited on flat metal surfaces and the local surface plasmon (LSP) excited by metalnanoparticles. It is known that fluorescence signals are enhanced by these two SPR-fields.On the other hand, fluorescence is quenched by the energy transfer to metal (FRET). Bothphenomena are controlled by the distance between dyes and metals, and the degree offluorescence enhancement is determined by the correlation. In this study, we determined thecondition to achieve the maximum fluorescence enhancement by adjusting the distance of ametal nanoparticle 2D sheet and a quantum dots 2D sheet by the use of $SiO_2$ spacer layers. The 2D sheets consisting of myristate-capped Ag nanoparticles (AgMy nanosheets) wereprepared at the air-water interface and transferred onto hydrophobized gold thin films basedon the Langmuir-Schaefer (LS) method [1]. The $SiO_2$ sputtered films with different thickness (0~100 nm) were deposited on the AgMy nanosheet as an insulator. TOPO-cappedCdSe/CdZnS/ZnS quantum dots (QDs, ${\lambda}Ex=638nm$) [2] were also transferred onto the $SiO_2$ films by the LS method. The layered structure is schematically shown in Fig. 1. The result of fluorescence measurement is shown in Fig. 2. Without the $SiO_2$ layer, the fluorescence intensity of the layered QD film was lower than that of the original QDs layer, i.e., the quenching by FRET was predominant. When the $SiO_2$ thickness was increased, the fluorescence intensity of the layered QD film was higher than that of the original QDs layer, i.e., the SPR enhancement was predominant. The fluorescence intensity was maximal at the $SiO_2$ thickness of 20 nm, particularly when the LSPR absorption wavelength (${\lambda}=480nm$) was utilized for the excitation. This plasmonic nanosheet can be integrated intogreen or bio-devices as the creation point ofenhanced LSPR field.

• Journal of the Optical Society of Korea
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• v.18 no.3
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• pp.274-282
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• 2014

In this study, closely spaced Au nanoparticles which are arranged in nanocluster (heptamer) configurations have been employed to design efficient plasmonic subwavelength devices to function at the telecommunication spectrum (${\lambda}$~1550 nm). Utilizing two kinds of nanoparticles, the optical properties of heptamer clusters composed of Au rod and shell particles that are oriented in triphenylene molecular fashion have been investigated numerically, and the cross-sectional profiles of the scattering and absorption of the optical power have been calculated based on a finite-difference time-domain (FDTD) method. Plasmon hybridization theory has been utilized as a theoretical approach to characterize the features and properties of the adjacent and mutual heptamer clusters. Using these given nanostructures, we designed a complex four-branch ($1{\times}4$) Y-shape splitter that is able to work at the near infrared region (NIR). This splitter divides and transmits the magnetic plasmon mode along the mutual heptamers arrays. Besides, as an important and crucial parameter, we studied the impact of arm spacing (offset distance) on the guiding and dividing of the magnetic plasmon resonance propagation and by calculating the ratio of transported power in both nanorod and nanoshell-based structures. Finally, we have presented the optimal structure, that is the four-branch Y-splitter based on shell heptamers which yields the power ratio of 23.9% at each branch, 4.4 ${\mu}m$ decaying length, and 1450 nm offset distance. These results pave the way toward the use of nanoparticles clusters in molecular fashions in designing various efficient devices that are able to be efficient at NIR.