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Methacrylamidohistidine in Affinity Ligands for Immobilized Metal-ion Affinity Chromatography of Human Serum Albumin

  • Odaba, Mehmet;Garipacan, Bora;Dede, Semir;Denizli, Adil
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.6
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    • pp.402-409
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    • 2001
  • Different bioligands carrying synthetic adsorbents have been reported in the literature for protein separation, We have developed a novel and new approach to obtain high protein ad-sorption capacity utilizing 2-methacrylamidohistidine(MAH) as a bioligand. MAH was synthe-sized by reacting methacrylocholride and histidine, Spherical beads with an average size of 150-200㎛ were obtained by the radical suspension polymerization of MAH and 2-hydrosyethyl-methacrylate(HEMA) conducted in an aqueous dispersion medium. p(HEMA-co-MAH) beads had a specific surface area of 17.6㎡/g . Synthesized MAH monomer was characterized by NMR. p(HEMA-co-MAH) beads were characterized by swelling test, FTIR and elemental analysis. Then Cu(II) ions were incorporated onto the beads and Cu(II) loading was found to be 0.96 mmol/g.These affinity beads with a swelling ration of 65% and containing, 1.6 mmol MAH/g were used in the adsorption/desorption of human serum albumin(HSA) from both aqueous solutions and hu-man serum. The adsorption of HSA onto p(HEM-co-MAH) was low(8.8 mg/g). Cu(II) chelation onto the beads significantly increased the HSA adsorption (56.3 mg/g). The maximum HSA ad-sorption ws observed at pH 8.0 Higher HSA adsorption was observed from human plasma(94.6 mgHSA/g) Adsorption of other serum proteins were obtained as 3.7 mg/g for fibrinogen and 8.5mg/g for γ-globulin. The total protein adsorption was determined as 107.1mg/g. Desorption of HSA was obtained using 0.1 M Tris/HCl buffer containing 0.5 M NaSCN, High desorption rations(up to 98% of the adsorbed HSA) were observed. It was possible to reuse Cu(II) chelated-p(HEMA-co-MAH) beads without significant decreases in the adsorption capacities.

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An Improved, Reliable and Practical Kinetic Assay for the Detection of Prekallikrein Activator in Blood Products

  • Shin, In-Soo;Shim, Yun-Bo;Hong, Choong-Man;Koh, Hyun-Chul;Lee, Seok-Ho;Hong, Seung-Hwa
    • Archives of Pharmacal Research
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    • v.25 no.4
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    • pp.505-510
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    • 2002
  • An improved kinetic assay for prekallikrein activator (PKA), a potential vasodilator, has been developed to be used as an indicator for quality control during production of human albumin preparations. It consists of two reaction stages. In the first stage, PKA and prekallikrein are incubated at $37^{\circ}C$ for 45 min to allow the transformation into kallikrein. Kallikrein, a serine protease, catalyzes the splitting of p-nitroaniline (pNA) from its substrate H-D-Pro-Phe-Arg-pNA(S-2302). The rate at which pNA is released was measured spectrophotometrically at 405 nm. Prekallikrein, a substrate of PKA was purified by DEAE ion-exchange chromatography and the major potential variations in the assay were optimized; pH 8.0 and 150 mM sodium chloride were chosen to give a proper ionic strength. Reaction times in the range of 10 to 360 min provided linear dose-response curves. The concentration of prekallikrein was adjusted to fall between 1:1 and 1:3 dilutions to generate a linear standard calibration curve. Under the optimized conditions, reproducibility was checked. In a precision test, the coefficient of variation (CV) stayed within ${\pm}4%$ and the dose-response curve showed a good correlation (${r^2}=0.999$). An accuracy test with an international standard of PKA afforded a mean recovery of 97.5%.

7Li MAS NMR studies of Li4P2O7 and LiFePO4 materials (LiFePO4와 Li4P2O77Li MAS NMR 특성 연구)

  • Han, Doug-Young;Park, Nam-Sin;Lee, Sang-Hyuk;Lee, Hak-Man;Kim, Chang-Sam
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.21 no.1
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    • pp.15-20
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    • 2011
  • [ $^7Li$ ]Magic Angle Spinning (MAS) NMR spectroscopy has been used to study the lithium local environments in $Li_4P_2O_7$ and$LiFePO_4$ materials. The purpose of this study was to know the structure of the solid electrolyte interphase (SEI) in lithium ion cells composed of $LiFePO_4$ as cathode material. $Li_4P_2O_7$ and $LiFePO_4$ were prepared by a solid-state reaction. The $^7Li$ MAS NMR experiments were carried out at variable temperatures in order to observe the local structure changes at the temperatures in $Li_4P_2O_7$ system. The $^7Li$ MAS NMR spectra of in $Li_4P_2O_7$ indicate that the lithium local environments in $Li_4P_2O_7$ were not changed in the temperature range between $27^{\circ}C$ and $97^{\circ}C$ Through this work, we confirmed that the small amount of $Li_4P_2O_7$ less than 5.0 wt% in $LiFePO_4$ could be clearly measured by the $^7Li$ MAS NMR spectroscopy at high spinning rate over than 11 kHz.

Physical Characteristics Comparison of Virtual Wedge Device with Physical Wedge (가상쐐기와 기존쐐기의 물리적 특성 비교)

  • Cho, Jung-Keun;Choi, Kye-Sook;Lim, Cheong-Hwan;Kim, Jeong-Koo;Jung, Hong-Ryang;Lee, Jung-Ok;Lee, Man-Goo
    • Journal of radiological science and technology
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    • v.24 no.2
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    • pp.49-52
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    • 2001
  • We compared the characteristics of Siemens virtual wedge device with physical wedges for clinical application. We investigated the characteristics of virtual and physical wedges for various wedge angles (15, 30, 45, and 60) using 6- and 15-MV photon beams. Wedge factors were measured in water using an ion chamber for various field sizes and depths. In case of virtual wedge device, as upper jaw moves during irradiation, wedge angles were estimated by accumulated doses. These measurements were performed at off-axis points perpendicular to the beam central axis in water for a $15\;cm\;{\times}\;20\;cm$ radiation field size at the depth of 10 cm. Surface doses without and with virtual or physical wedges were measured using a parallel plate ion chamber at surface. Field size was $15\;cm\;{\times}\;20\;cm$ and a polystyrene phantom was used. For various field sizes, virtual and physical wedge factors were changed by maximum 2.1% and 3.9%, respectively. For various depths, virtual and physical wedge factors were changed by maximum 1.9% and 2.9%, respectively. No major difference was found between the virtual and physical wedge angles and the difference was within 0.5. Surface dose with physical wedge was reduced by maximum 20% (x-ray beam : 6 MV, wedge angle : 45, SSD : 80 cm) relative to one with virtual wedge or without wedge. Comparison of the characteristics of Siemens virtual wedge device with physical wedges was performed. Depth dependence of virtual wedge factor was smaller than that of physical wedge factor. Virtual and physical wedge factors were nearly independent of field sizes. The accuracy of virtual and physical wedge angles was excellent. Surface dose was found to be reduced using a physical wedge.

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Elution characteristics of lime-based granular alkaline material and applicability of phosphorus crystallization processes (석회계 입상알칼리재의 용출특성과 이를 이용한 인 결정화공정의 적용성)

  • Chang, Hyang-Youn;Park, Na-Ri;Jang, Yeo-Ju;Ahn, Kwang-Ho;Lim, Hyun-Man;Kim, Weon-Jae
    • Journal of Korean Society of Water and Wastewater
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    • v.31 no.6
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    • pp.577-586
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    • 2017
  • One of the major sources causing eutrophication and algal blooms of lakes or streams is phosphorus which comes from point and nonpoint pollution sources. HAP (hydroxyapatite) crystallization using granular alkaline materials can achieve the decrease of phosphorus load from wastewater treatment plants and nonpoint pollution control facilities. In order to induce HAP crystal formation, continuous supply of calcium and hydroxyl ions is required. In this research, considering HAP crystallization, several types of lime-based granular alkaline materials were prepared, and the elution characteristics of calcium and hydroxyl ions of each were analyzed. Also, column tests were performed to verify phosphorus removal efficiencies of granular alkaline materials. Material_1 (gypsum+cement mixed material) achieved the highest pH values in the column tests consistently, also, Material_2 (gypsum+slag mixed material) and Material_3 (calcined limestone material) achieved over pH 9.0 for 240 hours (10 days) and proved the efficiencies of long-term ion supplier for HAP crystallization. In the column tests using Material_3, considerable pH increase and phosphorus removal were carried out according to each linear velocity and filtration depth. T-P removal efficiencies were 87.0, 84.0, 68.0% and those of PO4-P 100.0, 97.0, 80.0% for linear velocity of 1.0, 2.5, 5.0 m/hr respectively. Based on the column test results, the applicability of phosphorus removal processes for small-scale wastewater treatment plants and nonpoint pollution control facilities was found out.

Reference Dosimetry and Calibration of Glass Dosimeters for Cs-137 Gamma-rays (연구용 세슘-137 조사기에 대한 흡수선량 측정과 유리선량계 교정에 관한 연구)

  • Moon, Young Min;Rhee, Dong Joo;Kim, Jung Ki;Kang, Yeong-Rok;Lee, Man Woo;Lim, Heuijin;Jeong, Dong Hyeok
    • Progress in Medical Physics
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    • v.24 no.3
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    • pp.140-144
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    • 2013
  • In this research, the glass dosimeter was calibrated to measure the standard absorbed dose of the Cs-137 irradiator and absorbed dose in a biological sample. Absorbed dose in water for Cs-137 gamma ray was determined by the IAEA TRS-277 protocol. The PTW-TM30013 ion chamber and the PTW-TM41023 water phantom were utilized for measuring absorbed dose and the value was compared with the reading from DoseAce GD-302M glass dosimeter from Asahi Techno Glass Corporation for its calibration. The uncertainty of measurement ($1{\sigma}$) of the calibrated glass dosimeter was 2.7% and this result would be applied to improve the accuracy in measurement of absorbed dose in a biological sample.

Evaluation of Endogeneous Steroids Profile after Administration of Anabolic Steroids (근육 강화제 복용에 의한 내인성 스테로이드 프로필 평가)

  • Lee, Seon Hwa;Choi, Man Ho;Kim, Tae Wook;Chung, Bong Chul
    • Journal of the Korean Chemical Society
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    • v.41 no.8
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    • pp.406-413
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    • 1997
  • The endogenous steroids from human urine were simultaneously analyzed by selected ion monitoring method of GC/MS which is currently used for the doping procedure, together with anabolic steroids. The recovery range of this method was 72.33 %∼94.54% and the RSD values of precision and accuracy test were 1.43%∼10.86%, 0.96%∼9.98%, respectively. Using this method steroids profile was investigated in the urine of male volunteers after oral administration of nine anabolic steroids banned by IOC (International Olympic Committee). Urinary endogenous steroids level was varied specifically according to the excretion tendency of the metabolites of anabolic steroids.

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Patterns of Plasma Fatty Acids in Rat Models with Adenovirus Infection

  • Paik, Man-Jeong;Park, Ki-Ho;Park, Joong-Jean;Kim, Kyoung-Rae;Ahn, Young-Hwan;Shin, Gyu-Tae;Lee, Gwang
    • BMB Reports
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    • v.40 no.1
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    • pp.119-124
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    • 2007
  • Adenoviral vectors are among the most promising vectors available for human gene therapy. However, the use of recombinant adenoviral vectors, including replicationcompetent adenovirus (RCA), raises a variety of safety concerns in relation to the development of new therapies based on gene therapy. To examine how organic compounds change in rat plasma following the injection of adenovirus, $\beta$-galactosidase expressing recombinant adenovirus (designated rAdLacZ) or RCA, we investigated the content of fatty acids (FAs), which are important biochemical indicators in pathological conditions. Pattern recognition analysis on the level of FAs in rat plasma is described for the visual discrimination of adenovirus infection groups from normal controls. Plasma FAs from four control rats (normal group), and from four rats with rAdLacZ infection and six rats with RCA infection (the two abnormal groups), were examined by gas chromatography-mass spectrometry in selected ion monitoring modes as their tert-butyldimethylsilyl derivatives. In total, 20 FAs were positively detected and quantified. The results of the Student's t-test on the normal mean of two abnormal groups, the levels of three FAs (p<0.05) from rAdLacZ group and eleven FAs (p<0.05) from RCA group were significantly different. When star symbol plotting was applied to the group mean values of 20 FAs after normalization to the corresponding normal mean values, the resulting eicosagonal star patterns of the two infected groups were distorted into similar shapes, but were distinguishable from each other. Thus, these approaches will be useful for screening and monitoring of diagnostic markers for the effects of infection following the use of adenoviral vectors in gene therapy.

Review of Analytical and Assessment Techniques of Terminal Electron Accepting Processes (TEAPs) for Site Characterization and Natural Attenuation in Contaminated Subsurface Environments (오염 지중환경 특성화와 자연저감평가를 위한 말단전자수용과정(TEAPs) 분석 및 평가기술 소개)

  • Song, Yun Sun;Kim, Han-Suk;Kwon, Man Jae
    • Journal of Soil and Groundwater Environment
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    • v.25 no.2_spc
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    • pp.1-15
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    • 2020
  • Monitoring and assessing terminal electron accepting processes (TEAPs) are one of the most important steps to remediate contaminated sites via various in-situ techniques. TEAPs are a part of the microbial respiration reactions. Microorganisms gain energy from these reactions and reduces pollutants. Monitoring TEAPs enables us to predict degradability of contaminants and degradation rates. In many countries, TEAPs have been used for characterization of field sites and management of groundwater wells. For instance, US Environmental Protection Agency (EPA) provided strategies for groundwater quality and well management by applying TEAPs monitoring. Denmark has also constructed TEAPs map of local unit area to develop effective groundwater managing system, particularly to predict and assess nitrogen contamination. In case of Korea, although detailed soil survey and groundwater contamination assessment have been employed, site investigation guidelines using TEAPs have not been established yet. To better define TEAPs in subsurface environments, multiple indicators including ion concentrations, isotope compositions and contaminant degradation byproducts must be assessed. Furthermore, dissolved hydrogen concentrations are regarded as significant evidence of TEAPs occurring in subsurface environment. This review study introduces optimal sampling techniques of groundwater and dissolved hydrogen, and further discuss how to assess TEAPs in contaminated subsurface environments according to several contamination scenarios.

Mitigating Metal-dissolution in a High-voltage 15 wt% Si-Graphite‖Li-rich Layered Oxide Full-Cell Utilizing Fluorinated Dual-Additives

  • Kim, Jaeram;Kwak, Sehyun;Pham, Hieu Quang;Jo, Hyuntak;Jeon, Do-Man;Yang, A-Reum;Song, Seung-Wan
    • Journal of Electrochemical Science and Technology
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    • v.13 no.2
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    • pp.269-278
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    • 2022
  • Utilization of high-voltage electrolyte additive(s) at a small fraction is a cost-effective strategy for a good solid electrolyte interphase (SEI) formation and performance improvement of a lithium-rich layered oxide-based high-energy lithium-ion cell by avoiding the occurrence of metal-dissolution that is one of the failure modes. To mitigate metal-dissolution, we explored fluorinated dual-additives of fluoroethylene carbonate (FEC) and di(2,2,2-trifluoroethyl)carbonate (DFDEC) for building-up of a good SEI in a 4.7 V full-cell that consists of high-capacity silicon-graphite composite (15 wt% Si/C/CF/C-graphite) anode and Li1.13Mn0.463Ni0.203Co0.203O2 (LMNC) cathode. The full-cell including optimum fractions of dual-additives shows increased capacity to 228 mAhg-1 at 0.2C and improved performance from the one in the base electrolyte. Surface analysis results find that the SEI stabilization of LMNC cathode induced by dual-additives leads to a suppression of soluble Mn2+-O formation at cathode surface, mitigating metal-dissolution event and crack formation as well as structural degradation. The SEI and structure of Si/C/CF/C-graphite anode is also stabilized by the effects of dual-additives, contributing to performance improvement. The data give insight into a basic understanding of cathode-electrolyte and anode-electrolyte interfacial processes and cathode-anode interaction that are critical factors affecting full-cell performance.