• Title/Summary/Keyword: l biosensor

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A New Concept for Efficient Sensitivity Amplification of a QCM Based Immunosensor for TNF-α by Using Modified Magnetic Particles under Applied Magnetic Field

  • Bahk, Yeon-Kyoung;Kim, Hyung-Hoon;Park, Deog-Su;Chang, Seung-Cheol;Go, Jeung-Sang
    • Bulletin of the Korean Chemical Society
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    • v.32 no.12
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    • pp.4215-4220
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    • 2011
  • This study introduces a new concept for a simple, efficient and cheap sensitivity amplification of a Quartz Crystal Microbalance (QCM) based immunosensor system for the detection of tumor necrosis factor-alpha (TNF-${\alpha}$, TNF) by using an in-built magnetic system. The frequency shift due to the applied magnetic field was successfully observed on magnetic particles labeled detection antibodies, anti-human TNF-${\alpha}$, which were bound to the immunologically captured TNF-${\alpha}$ on the gold coated quartz crystals. In the present system, the magnitude of frequency shift depends on both the strength of magnetic field and the amount of target antigen applied. Significant signal amplification was observed when the additional built-in residual stress generated by the modified magnetic particles under the magnetic field applied. Used in conjunction with a sandwich type non-competitive immunoassay format, the lower detection limit was calculated to be 25 $ngmL^{-1}$ and showed good linearity up to TNF-${\alpha}$ concentrations as high as 2.0 ${\mu}gmL^{-1}$. The sensitivity, most importantly, was improved up to 4.3 times compared with the same QCM system which was used only an antigen-antibody binding without additional magnetic amplification.

A fiber optic surface plasmon resonance (SPR) sensorusing cyclic olefin copolymer (COC) polymer prism (Cyclic olefin copolymer (COC) 폴리머 프리즘을 사용한 광섬유 기반 표면 플라즈몬 공명 (SPR) 바이오 센서)

  • Yun, Sung-Sik;Lee, Soo-Hyun;Ahn, Chong-H.;Lee, Jong-Hyun
    • Journal of Sensor Science and Technology
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    • v.17 no.5
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    • pp.369-374
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    • 2008
  • A novel fiber optic surface plasmon resonance (SPR) sensor using cyclic olefin copolymer (COC) prism with the spectral modulation is presented. The SPR sensor chip is fabricated using the SU-8 photolithography, Ni-electroplating and COC injection molding process. The sidewall of the COC prism is partially deposited with Au/Cr (45/2.nm thickness) by e-beam evaporator, and the thermal bonding process is conducted for micro fluidic channels and optical fibers alignment. The SPR spectrum for a phosphate buffered saline (0.1.M PBS, pH.7.2) solution shows a distinctive dip at 1300.nm wavelength, which shifts toward longer wavelength with respect to the bovine serum albumin (BSA)concentrations. The sensitivity of the wavelength shift is $1.16\;nm{\cdot}{\mu}g^{-1}{\cdot}{\mu}l^{-1}$. From the wavelength of SPR dips, the refractive indices (RI) of the BSA solutions can be theoretically calculated using Kretchmann configuration, and the change rate of the RI was found to be $2.3{\times}10^{-5}RI{\cdot}{\mu}g^{-1}{\cdot}l^{-1}$. The realized fiber optic SPR sensor with a COC prism has clearly shown the feasibility of a new disposable, low cost and miniaturized SPR biosensor for biochemical molecular analyses.

Monitoring of Environmental Arsenic by Cultures of the Photosynthetic Bacterial Sensor Illuminated with a Near-Infrared Light Emitting Diode Array

  • Maeda, Isamu;Sakurai, Hirokazu;Yoshida, Kazuyuki;Siddiki, Mohammad Shohel Rana;Shimizu, Tokuo;Fukami, Motohiro;Ueda, Shunsaku
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1306-1311
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    • 2011
  • Recombinant Rhodopseudomonas palustris, harboring the carotenoid-metabolizing gene crtI (CrtIBS), and whose color changes from greenish yellow to red in response to inorganic As(III), was cultured in transparent microplate wells illuminated with a light emitting diode (LED) array. The cells were seen to grow better under near-infrared light, when compared with cells illuminated with blue or green LEDs. The absorbance ratio of 525 to 425 nm after cultivation for 24 h, which reflects red carotenoid accumulation, increased with an increase in As(III) concentrations. The detection limit of cultures illuminated with near-infrared LED was 5 ${\mu}g$/l, which was equivalent to that of cultures in test tubes illuminated with an incandescent lamp. A near-infrared LED array, in combination with a microplate, enabled the simultaneous handling of multiple cultures, including CrtIBS and a control strain, for normalization by the illumination of those with equal photon flux densities. Thus, the introduction of a near-infrared LED array to the assay is advantageous for the monitoring of arsenic in natural water samples that may contain a number of unknown factors and, therefore, need normalization of the reporter event.

A New Methodology of Measuring Water Toxicity using Sulfur Oxidizing Bacteria (황산화미생물을 이용한 새로운 수(水)중 생태독성탐지 방법)

  • Oh, Sang-Eun
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.6
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    • pp.555-562
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    • 2010
  • For the rapid and reliable detection of toxic compounds in water, a novel toxicity detection methodology based on sulfur-oxidizing bacteria (SOB) has been developed. The methodology exploits the ability of SOB to oxidize elemental sulfur to sulfuric acid in the presence of oxygen. The reaction results in an increase in electrical conductivity (EC) and a decrease in pH. Using a synthetic stream water (EC=0.12 mS/cm and pH=7.2), the baseline steady-state EC and pH values were 0.5~1.2 mS/cm and ~2.5 over 7 days of testing at HRT 30 minutes. When nitrite compounds were added to the system, the effluent EC decreased and the pH increased due to the inhibition of the SOB. Optimum HRT was 30 min and this HRT could be decresed by using smaller sulfur particles.

Real-time FRET imaging of cytosolic FAK signal on microwavy patterned-extracellular matrix (ECM) (미세파상 패턴 ECM 에서 세포질 FAK 신호의 실시간 FRET 이미징)

  • Suh, Jung-Soo;Jang, Yoon-Kwan;Kim, Tae-Jin
    • Journal of Biomedical Engineering Research
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    • v.40 no.1
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    • pp.1-6
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    • 2019
  • Human mesenchymal stem cells (hMSC) are multipotent stromal cells that have great potential to differentiate into a variety of cell types such as osteocytes, chondrocytes, and myocytes. Although there have been many studies on their clinical availability, little is known about how intracellular signals can be modulated by topographic features of the extracellular matrix (ECM). In this study, we investigated whether and how microwavy-patterned extracellular matrix (ECM) could affect the signaling activity of focal adhesion kinase (FAK), a key cellular adhesion protein. The fluorescence resonance energy transfer (FRET)-based FAK biosensor-transfected cells are incubated on microwavy-patterned surfaces and then platelet derived growth factor (PDGF) are treated to trigger FAK signals, followed by monitoring through live-cell FRET imaging in real time. As a result, we report that PDGF-induced FAK was highly activated in cells cultured on microwavy-patterned surface with L or M type, while inhibited by H type-patterned surface. In further studies, PDGF-induced FAK signals are regulated by functional support of actin filaments, microtubules, myosin-related proteins, suggesting that PDGF-induced FAK signals in hMSC upon microwavy surfaces are dependent on cytoskeleton (CSK)-actomyosin networks. Thus, our findings not only provide new insight on molecular mechanisms on how FAK signals can be regulated by distinct topographical cues of the ECM, but also may offer advantages in potential applications for regenerative medicine and tissue engineering.

In vitro Cytotoxicity Evaluation of Polydimethylsiloxane as a Biosensor Coating Material (바이오센서 코팅용 Polydimethylsiloxane의 생체외 세포독성 평가)

  • Park, Subeom;Lee, Jonghwan;Na, Kyunga;Jung, Jaeyeon;Kim, Myungjin;Park, Sungjae;Hyun, Jinho
    • Journal of Adhesion and Interface
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    • v.10 no.2
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    • pp.77-83
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    • 2009
  • PDMS was selected for a coating material of implantable biosensors and the cytotoxicity of extracts released from a polymer was evaluated using ISO 10993-5, Biological evaluation of medical devices-Part 5: Tests for in vitro cytotoxicity. Organo-tin was used as a positive control and a medium without serum was used as a negative control. Materials extract were prepared by incubating specimens in RPMI medium without serum ($125{\mu}L/cm^2$) for 24 h, 1 week and 6 weeks at $38^{\circ}C$. The evaluation of cytotoxicity was performed by two different methods : 1) seeding cells with extracts at the beginning 2) incubating extracts with cell sheets already formed on the plate. Both cell morphology and MTT numerical data were shown for the confirmation of cytotoxicity and cell spreading on the surface of PDMS.

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Effect of Bioluminescence Stimulating Agent of the Genetically Engineered Strain KG1206 on the Monitoring of the Petroleum Hydrocarbon Contaminated Groundwater Samples (발광유전자 재조합 균주 활성 촉진 조건이 석유계 탄화수소 오염지하수 모니터링에 미치는 영향)

  • Ko, Kyung-Seok;Kong, In-Chul
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.1
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    • pp.79-84
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    • 2008
  • This paper describes the application of bioluminescence stimulating agents on a genetically engineered microorganism, Pseudomonas putida mt-2 KG1206, to monitor toluene analogs using in groundwater samples from petroleum hydrocarbon contaminated sites. The maximum bioluminescent response with pure chemicals followed in the order: m-methyl benzyl alchohol > m-toluate > toluene > m-xylene > benzoate > p-xylene > o-xylene. Generally, the bioluminescence production of strain mixed with groundwater samples was dependent on the contaminated total inducer concentrations. However, few samples showed opposite results, where these phenomena may be caused by the complexicity of environmental samples. Two chemicals, SL(sodium lactate) and KNO$_3$, were tested to determine a better bioluminescence stimulant. Both chemicals stimulate the bioluminescence activity of strain KG1206, however, a slightly high bioluminescence was observed with nitrogen chemical. This selected stimulant was then tested on samples collected from contaminated groundwater samples. The bioluminescence activity of all samples mixed with the strain was stimulated with KNO$_3$ amendment. This suggests that the low bioluminescence activity exhibited by the environmental groundwater samples can be stimulated by amending the culture with a proper agent, such as nitrogen compound. These findings would be useful, especially, when strain was used to monitor the groundwater samples contaminated with low inducer contaminants. Overall, the results of this study found the ability of bioluminescence producing bacteria to biosensor a specific group of environmental contaminants, and suggest the potential for more efficient preliminary application of this engineered strain in a field-ready bioassay.