• Title/Summary/Keyword: Protein sensor

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QCM biosensor for the determination of haptoglobin (헵토글로빈 측정을 위한 수정미소저울 바이오센서 개발)

  • Kim, Sung-Il;Lee, Hyeon-Jeong;Park, Soon-Hyuk;Bhang, Hyun-Soo;Lee, Jin-Ah;Choi, Suk-Jung
    • Journal of Sensor Science and Technology
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    • v.16 no.2
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    • pp.132-141
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    • 2007
  • The purpose this research is to develop QCM (quartz crystal microbalance) biosensor for the determination of haptoglobin. Haptoglobin is an acute-phase protein with a hemoglobin-binding activity and has a potential to be used as a biomarker for infection or cancer. Haptoglobin level in milk has been used for the diagnosis of cow mastitis. In this study, anti-bovine haptoglobin antibody or bovine hemoglobin was chemically immobilized on the surface of the QCM, and the resulting sensor chips were tested for their response to samples containing haptoglobin at different concentrations. Concentration dependent frequency change was observed with both of the sensor chips. Especially, the sensor chip containing anti-bovine haptoglobin antibody showed sufficient sensitivity in the concentrations typically observed in the cows with mastitis.

v-Crk Induces Rac-dependent Membrane Ruffling and Cell Migration in CAS-deficient Embryonic Fibroblasts

  • Sung, Bong Hwan;Yeo, Myoung Gu;Oh, Hye Jin;Song, Woo Keun
    • Molecules and Cells
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    • v.25 no.1
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    • pp.131-137
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    • 2008
  • Crk-associated substrate (CAS) is a focal adhesion protein that is involved in integrin signaling and cell migration. CAS deficiency reduces the migration and spreading of cells, both of which are processes mediated by Rac activation. We examined the functions of v-Crk, the oncogene product of the CT10 virus p47gag-crk, which affects cell migration and spreading, membrane ruffling, and Rac activation in CAS-deficient mouse embryonic fibroblasts (CAS-/- MEFs). CAS-/- MEFs showed less spreading than did CAS+/+ MEFs, but spreading was recovered in mutant cells that expressed v-Crk (CAS-/-v-Crk MEF). We observed that the reduction in spreading was linked to the formation of membrane ruffles, which were accompanied by Rac activation. In CAS-/- MEFs, Rac activity was significantly reduced, and Rac was not localized to the membrane. In contrast, Rac was active and localized to the membrane in CAS-/-v-Crk MEFs. Lamellipodia protrusion and ruffle retraction velocities were both reduced in CAS-/- MEFs, but not in CAS-/-v-Crk MEFs. We also found that microinjection of anti-gag antibodies inhibited the migration of CAS-/-v-Crk MEFs. These findings indicate that v-Crk controls cell migration and membrane dynamics by activating Rac in CAS-deficient MEFs.

Effect of a Fermented Rice Protein Residue on the Taste Property of Yeast Extract (쌀단백질 잔사발효물이 효모추출물의 맛특성에 미치는 영향)

  • Park, Gang-Seok;Han, Gwi-Jung;Chung, Ha-Yull
    • Food Engineering Progress
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    • v.15 no.4
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    • pp.413-419
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    • 2011
  • For producing a high added-value natural seasoning ingredient, a yeast extract (Yx) was supplemented with a rice protein residue fermented with Bacillus licheniformis (Rfl) or with Bacillus subtilis (Rfs). A rice protein residue was obtained after enzymatic hydrolysis of rice protein which was used for preparing a yeast culture medium. Overall acceptabilities of the supplemented yeast extracts (YxRfl or YxRfs) were higher compared to pure yeast extract. Savory taste like umami was found to increase noticeably by adding a fermented rice protein residue to yeast extract, which was confirmed in taste sensor analysis and in sensory test. Beyond the presence of savory tasting amino acids such as Glu and Asp in a fermented rice protein residue, it is assumed that other soluble peptide fractions remained play an important role in enhancing taste of the supplemented yeast extracts. Thus, the yeast extract added with a fermented rice protein residue could be applied to manufacture a natural seasoning ingredient.

Role of AMP-Activated Protein Kinase (AMPK) in Smoking-Induced Lung Inflammation and Emphysema

  • Lee, Jae Seung;Park, Sun Joo;Cho, You Sook;Huh, Jin Won;Oh, Yeon-Mok;Lee, Sang-Do
    • Tuberculosis and Respiratory Diseases
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    • v.78 no.1
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    • pp.8-17
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    • 2015
  • Background: AMP-activated protein kinase (AMPK) not only functions as an intracellular energy sensor and regulator, but is also a general sensor of oxidative stress. Furthermore, there is recent evidence that it participates in limiting acute inflammatory reactions, apoptosis and cellular senescence. Thus, it may oppose the development of chronic obstructive pulmonary disease. Methods: To investigate the role of AMPK in cigarette smoke-induced lung inflammation and emphysema we first compared cigarette smoking and polyinosinic-polycytidylic acid [poly(I:C)]-induced lung inflammation and emphysema in $AMPK{\alpha}1$-deficient ($AMPK{\alpha}1$-HT) mice and wild-type mice of the same genetic background. We then investigated the role of AMPK in the induction of interleukin-8 (IL-8) by cigarette smoke extract (CSE) in A549 cells. Results: Cigarette smoking and poly(I:C)-induced lung inflammation and emphysema were elevated in $AMPK{\alpha}1$-HT compared to wild-type mice. CSE increased AMPK activation in a CSE concentration- and time-dependent manner. 5-Aminoimidazole-4-carboxamide-1-${\beta}$-4-ribofuranoside (AICAR), an AMPK activator, decreased CSE-induced IL-8 production while Compound C, an AMPK inhibitor, increased it, as did pretreatment with an $AMPK{\alpha}1$-specific small interfering RNA. Conclusion: $AMPK{\alpha}1$-deficient mice have increased susceptibility to lung inflammation and emphysema when exposed to cigarette smoke, and AMPK appears to reduce lung inflammation and emphysema by lowering IL-8 production.

Development of a Continuous High-Speed Single-Kernel Brown Rice Sorting Machine Based on Rice Protein Content

  • Natsuga, Motoyasu;Nakamura, Akitoshi;Kawano, Sumio
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.1616-1616
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    • 2001
  • To select kernels for breeding that have required constituent content from either naturally distributed samples or artificially mutated ones, it is necessary to process batch samples in a short time. The constituent content of single-kernel grains such as wheat and rice has been determined using conventional bench type NIR instruments; however, it takes a lot of time and effort. Shizuoka Seiki (Fukuroi-city, Japan) and NFRI (National Food Research Institute) of MAFF (Ministry of Agriculture, forestry and Fisheries of Japan) have jointly developed a continuous high-speed single-kernel brown rice sorting machine based on rice protein content. It consists of several sections such as a feeding mechanism, measuring unit, sorting mechanism and controlling PC. The feeding mechanism picks up single-kernel brown rice from the hopper (maximum of 5kg storage capacity) and sends it to the measuring unit. A spectrum of the brown rice is obtained in the measuring unit, which consists of a near-infrared array sensor. The brown rice is then sorted in the sorting mechanism based on its protein content estimated by the controlling PC. In the present study, measuring speed was approximately 500ms for the full spectrum range and overall sorting speed was approximately 2.8s for one kernel. Accuracy of estimation was approximately SEP=0.5% of dry matter protein content for nonglutinous rice.

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Easy Detection of Amyloid β-Protein Using Photo-Sensitive Field Effect

  • Kim, Kwan-Soo;Ju, Jong-Il;Song, Ki-Bong
    • Journal of Sensor Science and Technology
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    • v.21 no.5
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    • pp.339-344
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    • 2012
  • This article describes a novel method for the detection of amyloid-${\beta}$($A{\beta}$) peptide that utilizes a photo-sensitive field-effect transistor (p-FET). According to a recent study, $A{\beta}$ protein has been known to play a central role in the pathogenesis of Alzheimer's disease (AD). Accordingly, we investigated the variation of photo current generated from p-FET with and without intracellular magnetic beads conjugated with $A{\beta}$ peptides, which are placed on the p-FET sensing areas. The decrease of photo current was observed due to the presence of the magnetic beads on the channel region. Moreover, a similar characteristic was shown when the Raw 264 cells take in magnetic beads treated with $A{\beta}$ peptide. This means that it is possible to simply detect a certain protein using magnetic beads and a p-FET device. Therefore, in this paper, we suggest that our method could detect tiny amounts of $A{\beta}$ for early diagnosis of AD using the p-FET devices.

Development of Quantitative Vitellogenin ELISAs for Bullfrog (Rana catesbeiana) used in Endocrine Disrupter Screening (내분비계 장애물질 검색을 위한 효소면역측정법을 이용한 황소개구리 비텔로제닌 정량법 개발)

  • Lee Sang-Hoon;Kang Yun-Ju;Li Chun-Ri;Kim Andre;Jin Chun-Feng;Chung Kyu-Hyuck;Kim Dong-Kyoo;Park Nam-Gyu;Park Kwang-Sik;Kang Shin-Won;Park Jang-Su
    • Environmental Analysis Health and Toxicology
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    • v.21 no.2 s.53
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    • pp.147-151
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    • 2006
  • 난생 생물의 알 생성유도 단백질인 비텔로제닌(viteILogenin, VTG)을 성숙한 암컷 황소개구리 (Rana catesbeiana)혈청으로부터 음이온 교환 크로마토그래피를 이용하여 정제 하였으며 정제한 비텔로제닌을 BALB/c mice에 주사하여 폴리크로날 항체를 생산하였고 이것은 protein A column으로 정제 하였다. 이렇게 정제된 폴리크로날 항체를 이용하여 황소개구리 비탈로제닌 측정용 효소면역측정법을 개발하였으며 그 측정 범위는 $12{\sim}1,560ng/mL$였다. 또한 이 효소면역측정법을 평가하기 위해 성숙한 수컷 황소개구리를 청정지역과 폐수처리장 하류 하천에서 서식하는 황소개구리 혈액 속의 비텔로제닌을 측정하였다. 그 결과 폐수처리장 하류 하천에 서식하는 수컷 황소개구리 비텔로제닌이 청정지역보다 현저하게 높게 유도됨을 알 수 있었다.

Fluorescence Immunoassy of HDL and LDL Using Protein A LB Film

  • Choi, Jeong-Woo;Park, Jun-Hyo;Lee, Woo-Chang;Oh, Byung-Keun;Min, Jun-Hong;Lee, Won-Hong
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.979-985
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    • 2001
  • A fluorometric detection technique for HDL (High Density Lipoprotein) and LDL (Low Density Lipoprotein) was developed for application in a fiber-optic immunosensor using a protein A Langmuir-Blodgget (LB) film. For the fluorescence immunoassay, antibodies specific to HDL or LDL were imobilied on the protein A LB film, and a fluorescence amplification method was developed to overcome their weak fluorescence. The deposition of protein A using the LB technique was monitored using a surface pressure-are $({\pi}-A)$ curve, and the antibody immobilization of the protein A LB film was experimentally verified. The immobilized antibody was used to separate only HDL and LDL from a sample, then the fluorescence of he separated HDL or LDL was amplified. The amount of LDL or HDL was measured using the developed fiber optic fluorescence detection system. The optical properties resulting from the reaction of HDL or LDL with o-phtaldialdehyde, detection range, response time, and stability of the immunoassay were all investigated. The respective detection ranges for HDL and LDL were sufficient to diagnose the risk of coronary heart disease. The amplification step increased the sensitivity, while selective separation using the immobilized antibody led to linearity in the sensor signal. The regeneration of the antibody-immobilized substrate could produce a stable and reproducible immunosensor.

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

  • 김의락;최정우
    • KSBB Journal
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    • v.17 no.1
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    • pp.1-13
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    • 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.

Characterization of Protein Factor Regulating the Superoxide-Sensor SoxR in Escherichia coli

  • Koo, Mi-Sun;Rah, So-Yeon;Lee, Kang-Lok;Roe, Jung-Hye
    • Proceedings of the Korean Biophysical Society Conference
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    • 2001.06a
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    • pp.64-64
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    • 2001
  • Escherichia coli has developed soxRS regulon to defend against toxicity of superoxide radical. SoxR, superoxide sensor, is oxidized by superoxide-generating agents or nitric oxide and oxidized SoxR activates the transcription of soxS gene. In order to find out the trans-acting factors regulating SoxR activity in vivo, soxS::lacZ single copy operon fusion construct was prepared and random Tn10 insertional mutatons were performed.(omitted)

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