• Title/Summary/Keyword: SC1

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Production and Evaluation of Immunoreactivity of Poly Lysine-Tagged Single Chain Fragment Variable (ScFv) Lym-1 Antibody for Direct Conjugation to Fluorescence Dye (형광 물질 직접 표지를 위한 Poly Lysine 도입 Lym-1 단일사슬 항체의 제조 및 면역반응성 평가)

  • Jung, Jae-Ho;Choi, Tae-Hyun;Woo, Kwang-Sun;Chung, Wee-Sup;Kang, Joo-Hyun;Jeong, Su-Young;Choi, Chang-Woon;Lim, Sang-Moo;Cheon, Gi-Jeong
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.5
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    • pp.487-494
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    • 2009
  • Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.

Production of a Recombinant Anti-Human CD4 Single-Chain Variable-Fragment Antibody Using Phage Display Technology and Its Expression in Escherichia coli

  • Babaei, Arash;Zarkesh-Esfahani, Sayyed Hamid;Gharagozloo, Marjan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.529-535
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    • 2011
  • Single-chain variable fragment (scFv) is a fusion protein of the variable regions of the heavy (VH) and light (VL) chains of immunoglobulin, connected with a short linker peptide of 10 to about 20 amino acids. In this study, the scFv of a monoclonal antibody against the third domain of human CD4 was cloned from OKT4 hybridoma cells using the phage display technique and produced in E. coli. The expression, production, and purification of anti-CD4 scFv were tested using SDS-PAGE and Western blot, and the specificity of anti-CD4 scFv was examined using ELISA. A 31 kDa recombinant anti-CD4 scFv was expressed and produced in bacteria, which was confirmed by SDS-PAGE and Western blot assays. Sequence analysis proved the ScFv structure of the construct. It was able to bind to CD4 in quality ELISA assay. The canonical structure of anti-CD4 scFv antibody was obtained using the SWISS_MODEL bioinformatics tool for comparing with the scFv general structure. To the best of our knowledge, this is the first report for generating scFv against human CD4 antigen. Engineered anti-CD4 scFv could be used in immunological studies, including fluorochrome conjugation, bispecific antibody production, bifunctional protein synthesis, and other genetic engineering manipulations. Since the binding site of our product is domain 3 (D3) of the CD4 molecule and different from the CD4 immunological main domain, including D1 and D2, further studies are needed to evaluate the anti-CD4 scFv potential for diagnostic and therapeutic applications.

Effects of $UV/O_3$ and SC-1 Step in the HF Last Silicon Wafer Cleaning on the Properties of Gate Oxide (HF-last Cleaning에서 SC-1 step과 $UV/O_3$ step이 gate 산화막에 미치는 영향)

  • Choe, Hyeong-Bok;Ryu, Geun-Geol;Jeong, Sang-Don;Jeon, Hyeong-Tak
    • Korean Journal of Materials Research
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    • v.6 no.4
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    • pp.395-400
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    • 1996
  • 반도체 소자가 점점 고집적회되고 고성능화되면서 Si 기판 세정 방법은 그 중요성이 더욱 더 커지고 있다. 특히 ULSI급 소자에서는 세정 방법이 소자 생산수율 및 신뢰성에 큰 영향을 끼치고 있다. 본 연구에서는 HF-last 세정에 UV/O3과 SC-1 세정을 삽입하여 그 영향을 관찰하였다. 세정 방법은 HF-last 세정을 기본으로 split 1(piranha+HF), split 2(piranha+UV/O3+HF), split 3(piraha+SC-1+HF), split 4(piranha+(UV/O3+HF) x3회 반복)의 4가지 세정 방법으로 나누어 실험하였다. 세정을 마친 Si 기판은 Total X-Ray Fluorescence Spectroscopy(AFM)을 사용하여 표면거칠기를 측정하였다. 또한 세정류량을 측정하고, Atomic Force Microscopy(AFM)을 사용하여 표면거칠기를 측정하였다. 또한 세정후 250$\AA$의 gate 산화막을 성장시켜 전기적 특성을 측정하였다. UV/O3을 삽입한 split 2와 split 4세정방법이 물리적, 전기적 특성에서 우수한 특성을 나타냈고, SC-1을 삽입한 split 3세정 방법이 표준세정인 split 1세정 방법보다 우수하지 못한 결과를 나타냈다.

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The Quality characteristics of sponge cake added with the ingredients of Sagoonja-Tang (사군자탕(四君子湯) 재료를 첨가한 스폰지 케이크의 품질특성)

  • Nam, Hae-Young
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.4
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    • pp.1066-1075
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    • 2017
  • This study was carried out to determine the functional works and basic ingredients of Sagoonja-Tang on sponge cake. The experimental groups consisted of 6% mixtures of five (Ed-confirm the number) kinds powders : S1 for Sagoonja-Tang powder, S2 for Panax ginseng, S3 for Poria cocos, Koidz, S4 for Atractylodes macrocephala, and S5 for Glycyrrhiza uralensis Fisch. The volume of sponge cake somewhat decreased after adding the ingredients, and the volume differences in order from highest to lowest, were control>S1>S3>S2>S4>S5. In the microbiological quality test, viable cell counts were high in control groups of sponge cake, and the numbers of viable cell for control sponge cake reached to $9{\times}10^7CFU/g$ on the 7th day of storage, and decreased to $2.5{\times}10^5CFU/g$ on the 10th day of storage. All sponge cakes added with ingredient's powder showed pretty low viable cell counts. Especially, the group S5 showed the lowest counts of $1.2{\times}10^2CFU/g$ on the 7th day of storing. In the texture analyses of sponge cake, All groups showed higher degree of hardness, gumminess and chewiness than SC. The antioxidative activity of the Sagoonja-Tang's ingredients was measured. The POV value measured was S5>S4>S3>S2 >S1>control in order of highest to lowest. In sensory test, the overall acceptability of sponge cake was from highest to lowest S3>SC S2>S1>S5>S4.

High Temperature Deformation Behavior of Sc Added Al-7.7wt%Zn-2.0wt%Mg-1.9wt%Cu Alloy (Sc을 첨가한 Al-7.7wt%Zn-2.0wt%Mg-1.9wt%Cu합금의 고온 변형거동)

  • Woo, Kee-Do;Ryu, Yong-Seok;Kim, Sug-Won;Deliang Zhang
    • Korean Journal of Materials Research
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    • v.13 no.12
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    • pp.819-824
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    • 2003
  • The Al-7.7Zn-2.0Mg-1.9Cu-0.1Zr-0.1Sc alloy exhibited excellent elongation by the new thermomechanical treatment (TMT) process; solution treatment and furnace cooling\longrightarrowhot and cold rolling and then annealing for short time. Tensile test at high temperature from 430 to $500^{\circ}C$ has been performed with various strain rates using for the Al-7.7Zn-2.0Mg-1.9Cu-0.1Zr-0.1Sc alloy obtained by the TMT process. The elongation of the Al-7.7Zn-2.0Mg-1.9Cu-0.1Zr-0.1Sc was 550% tensile tested at $470^{\circ}C$ temperature and 2.2 $\times$ $10^{-3}$ $s^{-1}$ strain rate. The m value of Al-7.7Zn-2.0Mg-1.9Cu-0.1Zr-0.1Sc alloy deformed 85% increased from 0.33 to 0.46 with increasing total elongation. This new TMT process was very simple and easy to make the sheets in the company.

Effect of a Hot Water Extract of Sparasis Crispa on the Expression of Tight Junction-Associated Genes in HaCaT Cells (꽃송이버섯 열수추출물이 HaCaT의 세포 연접 관련 유전자의 발현에 대한 영향)

  • Han, Hyo-Sang
    • Journal of The Korean Society of Integrative Medicine
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    • v.9 no.2
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    • pp.83-92
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    • 2021
  • Purpose : Keratinocytes are the main cellular components involved in wound healing during re-epithelization and inflammation. Dysfunction of tight junction (TJ) adhesions is a major feature in the pathogenesis of various diseases. The purpose of this study was to identify the various effects of a Sparassis crispa water extract (SC) on HaCaT cells and to investigate whether these effects might be applicable to human skin. Methods : We investigated the effectiveness of SC on cell HaCaT viability using MTS. The antioxidant effect of SC was analyzed by comparing the effectiveness of ABTS to that of the well-known antioxidant resveratrol. Reverse-transcription quantitative polymerase chain reaction (qRT-PCR) is the most widely applied method Quantitative RT-PCR analysis has shown that SC in HaCaT cells affects mRNA expression of tight-junction genes associated with skin moisturization. In addition, Wound healing is one of the most complex processes in the human body. It involves the spatial and temporal synchronization of a variety of cell types with distinct roles in the phases of hemostasis, inflammation, growth, re-epithelialization, and remodeling. wound healing analysis demonstrated altered cell migration in SC-treated HaCaT cells. Results : MTS analysis in HaCaT cells was found to be more cytotoxic in SC at a concentration of 0.5 mg/㎖. Compared to 100 µM resveratrol, 4 mg/㎖ SC exhibited similar or superior antioxidant effects. SC treatment in HaCaT cells reduced levels of claudin 1, claudin 3, claudin 4, claudin 6, claudin 7, claudin 8, ZO-1, ZO-2, JAM-A, occludin, and Tricellulin mRNA expression by about 1.13 times. Wound healing analysis demonstrated altered cell migration in SC-treated HaCaT cells and HaCaT cell migration was also reduced to 73.2 % by SC treatment. Conclusion : SC, which acts as an antioxidant, reduces oxidative stress and prevents aging of the skin. Further research is needed to address the effects of SC on human skin given the observed alteration of mRNA expression of tight-junction genes and the decreased the cell migration of HaCaT cells.

Pressure Sensing Properties of Al1-xScxN Thin Films Sputtered at Room Temperature (상온에서 성막한 고감도의 Al1-xScxN 박막의 압력 감지 특성)

  • Seok, Hye-Won;Kim, Sei-Ki;Kang, Yang-Koo;Lee, Young-Jin;Hong, Yeon-Woo;Ju, Byeong-Kwon
    • Journal of Sensor Science and Technology
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    • v.23 no.6
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    • pp.420-424
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    • 2014
  • Aluminum-scandium nitride ($Al_{1-x}Sc_xN$) thin films with a TiN buffer layer have been fabricated on SUS430 substrate by RF reactive magnetron sputtering at room temperature under 50% $N_2$/Ar. The effect of Sc-doping on the structure and piezoelectric properties of AlN films has been investigated using SEM, XRD, surface profiler and pressure-voltage measurements. The as-deposited AlN films showed polycrystalline phase, and the Sc-doped AlN film, the peak of AlN (002) plane and the crystallinity became very strong. With Sc-doping, the crystal size of AlN film was grown from ~20 nm to ~100 nm. The output signal voltage of AlN sensor showed a linear behavior between 15~65 mV, and output signal voltage of Sc-doped AlN sensor was increased over 7 times. The pressure-sensing sensitivity of AlN film was calculated about 10.6mV/MPa, and $Al_{0.88}Sc_{0.12}N$ film was calculated about 76 mV/MPa.