• Title/Summary/Keyword: ProteinChip

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Automated Protein-Expression Profiling System using Crude Protein Direct Blotting Method

  • Kobayashi, Hironori;Torikoshi, Yasuhiro;Kawasaki, Yuko;Ishihara, Hideki;Mizumoto, Hiroshi
    • 제어로봇시스템학회:학술대회논문집
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    • 2003.10a
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    • pp.2356-2361
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    • 2003
  • Proteome research in the medical field is expected to accelerate the understanding of disease mechanism, and to create new diagnostic concept. For protein profiling, this paper proposes a new methodology named CPDIB (Crude Protein Direct Blotting). In the CPDIB procedure, crude protein sample is directly immobilized on a membrane and the expression of protein molecules in the sample are analyzed quantitatively by using a special device called ImmobiChip, where the membrane is used as a field of the immune reaction. The over-all structure of the ImmobiChip is based on the conventional Slot blot device. Mechanical improvement in the air-tightness of the case holding the membrane realizes the direct blotting and results in high performance of stability in the immune reaction. In the measurement of multiple proteins, a dispensing robot is used for increasing the efficiency of handling of liquid. Cooperation of the dispensing robot with the ImmobiChip for immobilizing proteins realizes automated and stable performance of the CPDIB procedure. This paper shows the evaluation of the air-tightness of the ImmobiChip, the ability of analyzing proteins using the CPDIB procedure and the performance of the automated equipment.

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Nanoscale Fabrication of Biomolecular Layer and Its Application to Biodevices

  • Park, Jeong-Woo;Nam, Yun-Suk;Masamichi Fujihira
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.2
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    • pp.76-85
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    • 2004
  • Biodevices composed of biomolecular layer have been developed in various fields such as medical diagnosis, pharmaceutical screening, electronic device, photonic device, environmental pollution detection device, and etc. The biomolecules such as protein, DNA and pigment, and cells have been used to construct the biodevices such as biomolecular diode, biostorage device, bioelectroluminescence device, protein chip, DNA chip, and cell chip. Substantial interest has focused upon thin film fabrication or the formation of biomaterials mono- or multi-layers on the solid surfaces to construct the biodevices. Based on the development of nanotechnology, nanoscale fabrication technology for biofilm has been emerged and applied to biodevices due to the various advantages such as high density immobilization and orientation control of immoblized biomolecules. This review described the nanoscale fabrication of biomolecular film and its application to bioelectronic devices and biochips.

Immunoassay of haptoglobin and transferrin with proteinG-containing QCM sensor chip and unpurified antiserum (Protein G를 포함하는 수정미소저울 센서 칩과 정제되지 않은 항혈청을 이용한 헵토글로빈과 트랜스페린의 면역분석)

  • Ha, In-Young;Choi, Suk-Jung
    • Journal of Sensor Science and Technology
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    • v.17 no.5
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    • pp.380-386
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    • 2008
  • Quartz crystal microbalance immunosensor has a capacity to perform a label-free and real time detection of a trace amount of analyte through the specific interaction between antibody and antigen. However, immobilization of antibody molecules on the sensor surface is a troublesome procedure for researchers who are not experienced in chemistry. Protein G has a specific affinity to antibody and would serve as a capturing agent for antibody when immobilized on the sensor surface. In this work, we prepared a protein G sensor chip by immobilizing protein G on the surface of quartz crystal microbalance and examined its capability to detect human haptoglobin or human transferrin with unpurified corresponding antiserum. Specific and dose dependent response was observed when the protein G chip was used for detection of antigens after saturated with antiserum. We also verified several advantageous aspects of the protein G chip such as improved flexibility and sensitivity.

Effect of Solvent on the Surface of Protein Chip Plate (단백질 칩 기판의 표면에 미치는 용매 효과)

  • 현준원;윤미영;안상민;노승정;허영덕;박헌용;송예신;피재호;김경례
    • Journal of the Korean institute of surface engineering
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    • v.37 no.2
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    • pp.76-79
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    • 2004
  • Nickel chloride coated protein chip plate was developed by using a spin coating method. The ability of histidine tagged protein adsorption was investigated at various solvents. The surface of plate has a large aggregated nickel complex with high density in water. However, the surface of plate has a very small size of aggregated nickel complex with low density in isopropanol. The ability of protein adsorption decreased as increasing the size of alkyl chain in various alcohol solvents. The mechanism on the ability of protein adsorption at the plate surface is discussed.

Effects of Scratching on the Surface of Protein Chip Plates (단백질 칩 기판의 표면 스크래칭 효과)

  • Hyun, June-Won;Hwang, Jeong-Il
    • Journal of the Korean institute of surface engineering
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    • v.40 no.2
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    • pp.98-102
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    • 2007
  • [ $NiCl_2$ ] and poly-L-lysine coated protein chip plates have been fabricated using a spin coating system. Water has been used as solvent and scratching effects on glass slides and ITO have been investigated. We also observed the surface properties of $NiCl_2$ and poly-L-lysine coated slides by using PSA(Particle size analyzer) and AFM(Atomic force microscope). The AFM results imply that the surface patterns created in the spin coating system determine the protein adsorption. Adsorption of histidine-tagged KRS proteins immobilized on glass slides and ITO was analyzed using a BAS image system. The results suggest that the scratching effect was increased ability of protein adsorption.

New role of E3 ubiquitin ligase in the regulation of necroptosis

  • Seo, Jinho;Lee, Eun-Woo;Song, Jaewhan
    • BMB Reports
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    • v.49 no.5
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    • pp.247-248
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    • 2016
  • Necroptosis is a well-known form of caspase-independent cell death. Necroptosis can be triggered by various extrinsic stimuli, including death ligands in the presence of receptorinteracting protein kinase 3 (RIPK3), a key mediator of necroptosis induction. Our recent studies have revealed that C-terminus HSC-70 interacting protein (CHIP), an E3 ligase, can function as an inhibitor of necroptosis. CHIP−/− mouse embryonic fibroblast showed higher sensitivity to necrotic stimuli than wild-type mouse embryonic fibroblast cells. Deleterious effects of CHIP knockout MEFs were retrieved by RIPK3 depletion. We found that CHIP negatively regulated RIPK3 and RIPK1 by ubiquitylation- and lysosome- dependent degradation. In addition, CHIP−/− mice showed postnatal lethality with intestinal defects that could be rescued by crossing with RIPK3−/− mice. These results suggest that CHIP is a negative regulator of RIPK1 and RIPK3, thus inhibiting necroptosis.

High Throughput Screening on Angiogenesis Inhibitor and Promoter of Medicinal Plants using a Protein Microarray Chip

  • In, Dong-Su;Lee, Min-Su;Bang, Kyong-Hwan;Kim, Ok-Tae;Hyun, Dong-Yun;Ahn, Young-Sup;Cha, Seon-Woo;Seong, Nak-Sul;Kim, Eung-Youn;Shin, Yoo-Soo;Kang, In-Cheol
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.2
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    • pp.89-94
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    • 2007
  • The effects of angiogenesis inhibitor from the extract libraries of Korean and Chinese medicinal plants were investigated using a protein microarray chip. Protein chip was constructed by immobilization of integrin ${\alpha}_5{\beta}_1$ on protein chip base plates and employed far screening active extracts that inhibit the integrin-fibronectin interaction from the extract libraries. The 100 extracts of medicinal plants were obtained from extract bank of National Institute of Crop Science, RDA. The 14 extracts among 100 extract libraries were shown efficient inhibition activity for the interaction between integrin-fibronectin. The medicinal plants of 14 extracts were Vitex negundo var. incisa (Lam.) C.B. Clarke, Epimedium koreanum Nakai, Cedrela sinensis A. Juss, Ipomea aquatica Forsk, Schisandra chinensis Baill, Pulsatilla koreana Nakai, Paeonia lactiflora Pall. var.hortensis Makino, Oenothera odorata, Allium chinense, Allium victorialis var. platyphyllum MAKINO, Polygonatum odoratum Druce var. pluriflorum Ohwi, Hosta lancifolia, Agrimonia pilosa L. var. japonica Nakai and Potentilla chinensis SER. The Paeonia lactiflora, Oenothera, and Agrimonia pilosa from these 14 extracts libraries were shown strong inhibition activity of integrin ${\alpha}_5{\beta}_1$.

Quantitative and Rapid Analysis of Transglutaminase Activity Using Protein Arrays in Mammalian Cells

  • Kwon, Mi-Hye;Jung, Jae-Wan;Jung, Se-Hui;Park, Jin-Young;Kim, Young-Myeong;Ha, Kwon-Soo
    • Molecules and Cells
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    • v.27 no.3
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    • pp.337-343
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    • 2009
  • We developed a novel on-chip activity assay using protein arrays for quantitative and rapid analysis of transglutaminase activity in mammalian cells. Transglutaminases are a family of $Ca^{2+}$-dependent enzymes involved in cell regulation as well as human diseases such as neurodegenerative disorders, inflammatory diseases and tumor progression. We fabricated the protein arrays by immobilizing N,N'-dimethylcasein (a substrate) on the amine surface of the arrays. We initiated transamidating reaction on the protein arrays and determined the transglutaminase activity by analyzing the fluorescence intensity of biotinylated casein. The on-chip transglutaminase activity assay was proved to be much more sensitive than the $[^3H]putrescine$-incorporation assay. We successfully applied the on-chip assay to a rapid and quantitative analysis of the transglutaminase activity in all-trans retinoic acid-treated NIH 3T3 and SH-SY5Y cells. In addition, the on-chip transglutaminase activity assay was sufficiently sensitive to determine the transglutaminase activity in eleven mammalian cell lines. Thus, this novel on-chip transglutaminase activity assay was confirmed to be a sensitive and high-throughput approach to investigating the roles of transglutaminase in cellular signaling, and, moreover, it is likely to have a strong potential for monitoring human diseases.

Miniature Fluorescence Detection System for Protein Chips by Prism (프리즘을 이용한 소형 단백질칩 분석 형광측정 시스템 개발)

  • Choi, Jae-Ho;Kim, Ho-Seong;Lee, Kook-Nyung;Kim, Eun-Mi;Kim, Yong-Kweon;Kim, Byung-Gee
    • Proceedings of the KIEE Conference
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    • 2004.07c
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    • pp.2040-2042
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    • 2004
  • This paper presents a miniature optical system for the fluorescence detection of the patterned protein chip. The patterned protein chip was fabricated using MEMS process. The fluorescence from the patterned protein chip was measured while varying the concentration of the BSA. The fluorescence light is separated spatially from the excitation beam using mini-size prism to increase SNR (Signal-to-Noise Ratio). The combination of prism and mirrors can convert the excitation light from the laser diode to uniform illumination on the specimen. We believe that the proposed system for fluorescence detection can be applied to rea1ization of point-of-care.

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