• Title/Summary/Keyword: Suspension stability

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Virus-like Particles Containing Cytokine Plasmid DNA (사이토카인 유전자 함유 바이러스 유사입자의 제조)

  • Oh, Yu-Kyoung;Son, Tae-Jong;Sin, Kwang-Sook;Kang, Min-Jeong;Kim, Jung-Mogg;Kim, Nam-Keun;Ko, Jung-Jae;Kim, Chong-Kook
    • Journal of Pharmaceutical Investigation
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    • v.31 no.3
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    • pp.185-190
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    • 2001
  • Human papillomavirus (HPV) infection is known to cause cervical cancers. Human papillomavirus-like particles (VLP) have been studied as preventive vaccines of cervical cancers. To develop VLP as a therapeutic gene carrier, we studied the method to encapsulate cytokine genes in virus-like particles. HPV type 16 capsid L1 genes were amplified by polymerase chain reaction and cloned into T vector. L1 gene was then inserted into baculovirus transfer vector. The clone of baculovirus encoding L1 gene was isolated and used to express L1 protein in Sf 21 insect cells. VLP were purified by CsCl density gradient and ultracentrifugation. VLP were disassembled to capsomer units by treatment of a reducing agent. Given that interleukin-2 (IL-2) genes have been used in anticancer gene therapy and as a molecular adjuvant, IL-2 cytokine plasmids were chosen as a model gene. IL-2 plasmids were incubated with the disassembled capsomer suspension. To reassemble the particles, the mixture of capsomers and cytokine plasmids was dialyzed. The disassembly and reassembly of VLP were confirmed by transmission electron microscopy. The entrapment of cytokine plasmids in reassembled VLP was tested by the stability of plasmids against DNase I. After treatment of reassembled virus-like particles with DNase I, discrete IL-2 DNA band was observed. Our results indicate that IL-2 cytokine plasmid (3.5 kb size) can be encapsulated in the virus-like particles, suggesting the potential of VLP as a gene delivery system. Moreover, VLP containing the adjuvant cytokine plasmids might function as more effective subunit vaccines.

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Dispersion Characteristics of Natural Crystalline Graphite Powders by Surface Modification (표면개질에 의한 인상흑연 분체의 분산특성 연구)

  • Kim, Byeong-Gon;Choe, Sang-Geun;Jeong, Heon-Saeng;Han, Sang-Geun;Lee, Jae-Jang
    • Korean Journal of Materials Research
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    • v.11 no.8
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    • pp.679-684
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    • 2001
  • The surface of natural graphite has not only good electrical conductivities and lubrication properties but also has strong hydrophobicity. There are no functional groups and chemical properties on it. It is difficult to join with any other ions and to disperse in aqueous system. In order to increase dispersion ability throughout modification of surface property, it is necessary to let graphite have some function on its surface by the adsorption of surfactant molecules. In this study, using zeta potential adsorbed surfactant molecules(ABDM) on graphite surface and its surface Properties turn hydrophobic into hydrophilic. The dispersing mechanism of graphite particles in aqueous system has been explained using the DLVO theory, It is concluded that the high dispersable graphite suspension of which dispersing stability$(T_{1/2})$ is 44.5 hours at pH 10 and 22.5mV zeta potential can be produced.

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Enhanced Production of hGM-CSF by Immobilized Transgenic Plant Cell Cultures (형질전환된 식물세포에서 고정화 방법을 통한 hCM-CSF의 생산성 증대 연구)

  • Noha, Yun-Sook;Nama, Hyung-Jin;Choi, Hong-Yeol;Tak, Sa-Ra;Kim, Dong-Il
    • KSBB Journal
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    • v.30 no.2
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    • pp.82-90
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    • 2015
  • Plant cell immobilization can protect plant cells from shear forces and increase the stability of gene. An additional advantage of immobilization is the easiness for performing continuous culture with cell recycling. Therefore plant cell immobilization can overcome the limitations of plant cell applications. In addition, target protein should be selected from pharmaceutical proteins to get rid of low expression level problem. The enhanced production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was investigated in immobilized Nicotiana tabacum suspension cell cultures. When the cells were immobilized in polyurethane foam, specific production of hGM-CSF was higher than that in alginate bead immobilization. Optimum continuous culture condition was the addition of 60 g/L sucrose in growth media with exchanging media every 6 day. Under the same condition, specific hGM-CSF production was 7 times higher in a 500-mL spinner flask than that in 100-mL Erlenmeyer flasks. Therefore, development of an effective immobilization process would be possible when the advantage of easy cell recycling was used. Consequently, enhanced production of target proteins could be possible in immobilized continuous cultures when the advantages of immobilization were applied.

In situ Recovery of hCTLA4Ig from Suspension Cell Cultures of Oryza sativa (형질전환 벼 현탁세포 배양에서 hCTLA4Ig의 in situ 회수)

  • Choi, Hong-Yeol;Cheon, Su-Hwan;Kwon, Jun-Young;Yun, Boreum;Hong, Seok-Mi;Kim, Sun-Dal;Kim, Dong-Il
    • KSBB Journal
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    • v.31 no.4
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    • pp.284-290
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    • 2016
  • In this research, recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced by transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenic plant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promoter system which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell death promotes the release of protease which degrades the target proteins. The protein stability and productivity can be significantly influenced by proteolysis activity. Therefore, development of new strategies are necessary for the in situ recovery of target proteins from cell culture media. In this study, in situ recovery was performed by various strategies. Direct addition of Protein A resin with nylon bag leads to cell death by increased shear stress and decrease in production of hCTLA4Ig by protease. Medium exchange through modified flask could recover hCTLA4Ig with high cell viability and low protease activity, on the other hand, the productivity was lower than that of control. When in situ recovery was conducted at day 7 after induction in air-lift bioreactor, 1.94-fold of hCTLA4Ig could be recovered compared to control culture without in situ recovery. Consequently, in situ recovery of hCTLA4Ig from transgenic rice cell culture could enhance productivity significantly and prevent degradation of target proteins effectively.

Dynamic Behavior of Direct Fixation Track on Yeongjong Grand Bridge (영종대교 강직결 궤도구조의 동적거동에 관한 연구)

  • Choi, Jung-Youl;Lee, Kyu-Yong;Chung, Jee-Seung;Ahn, Dae-Hee;Kim, Soo-Hyung
    • The Journal of the Convergence on Culture Technology
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    • v.6 no.3
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    • pp.443-448
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    • 2020
  • In this study, field measurements were performed to analyze the effects of train types (AREX, AREX Express, KTX, KTX-Sancheon) and train speeds on the dynamic behavior of the direct fixation track structure on Yeongjong grand bridge by bridge type (truss bridge, suspension bridge). Based on field measurement results, the track impact factor and train running stability (coefficient of derailment, Rate of wheel load reduction, lateral displacement of rail head) are compared with domestic and foreign standards and regulations to influence the dynamic behavior of direct fixation track. As a result, the differences in the dynamic behavior of the direct fixation tracks by the type of bridges of Yeongjong bridge are not significant, but it was analyzed that these were more directly affected by the magnitude of the train load. Therefore, it is necessary to establish the reinforcement plan of the direct fixation track structure on Yeongjong grand bridge in consideration of the increase of the track impact factor and dynamic track force.

Study on the Epoxy/BaTiO$_3$Embedded Capacitor Films for PWB Applications (인쇄회로기판 용 Epoxy/BaTiO$_3$내장형 커패시터 필름에 관한 연구)

  • 조성동;이주연;백경욱
    • Journal of the Microelectronics and Packaging Society
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    • v.8 no.4
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    • pp.59-65
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    • 2001
  • Epoxy/$BaTiO_3$composite capacitor films with excellent stability at room temperature, uniform thickness, and electrical properties over a large area ware successfully fabricated. The composite capacitor films with good film formation capability and easy process ability were made from epoxy resin developed for ACF as a matrix and two kinds of $BaTiO_3$powders as fillers to increase the dielectric constant of the composite film. The crystal structure of the powders and its effects on dielectric constant of the films were investigated by X-ray diffraction (XRD). And the optimum amount of dispersant, phosphate ester, was determined by viscosity measurement of suspension. DSC and dielectric property tests were conducted to decide the right curing temperature and the optimum amount of the curing agent. As a result, the capacitors of 7 $\mu \textrm{m}$ thick film with 10 nF/$\textrm{cm}^2$ and low leakage current were successfully demonstrated.

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A Numerical Study on Shear Behavior of the Interface between Blasted Rock and Concrete (발파 암반-콘크리트 경계면에서의 전단거동특성에 대한 수치해석적 연구)

  • Min, Gyeong-Jo;Ko, Young-Hun;Fukuda, Daisuke;Oh, Se-Wook;Kim, Jeong-Gyu;Chung, Moon-Kyung;Cho, Sang-Ho
    • Explosives and Blasting
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    • v.37 no.4
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    • pp.26-35
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    • 2019
  • In designing a gravity-type anchorage of earth-anchored suspension bridge, the contact friction between a blasted rock mass and the concrete anchorage plays a key role in the stability of the entire anchorage. Therefore, it is vital to understand the shear behavior of the interface between the blasted rock mass and concrete. In this study, a portable 3D LiDAR scanner was utilized to scan the blasted bottom surfaces, and rock surface roughness was quantitatively analyzed from the scanned profiles to apply to 3D FEM modelling. In addition, based on the 3D FEM model, a three-dimensional dynamic fracture process analysis (DFPA-3D) technique was applied to study on the shear behavior of the interface between blasted rock and concrete through direct shear tests, which was analyzed under constant normal load (CNL). The effects of normal stress and the joint roughness on shear failure behavior are also analyzed.

Studies on Preparation of Dysprosium-165 Metallic Macroaggregates for the Treatment of Rheumatoid Arthritis (류마티스 관절염 치료용 디스프로슘-165금속 응집입자($^{165}Dy-MA$)의 제조에 관한 연구)

  • Park, Kyung-Bae;Kim, Jae-Rok
    • The Korean Journal of Nuclear Medicine
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    • v.28 no.2
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    • pp.227-233
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    • 1994
  • Irradiation of 20mg of natural $Dy(NO_3)_3$ in a neutron flux of $2{\times}10^{13}n/cm^2$ sec for 4 hours gave 5.76 Ci of $^{165}Dy$(specific activity, 610mCi/mg Dy) with high radionuclidic purity (>99.9 %). $^{165}Dy-MA$ was prepared in a quantitative yield by reacting the aqueous solution of $^{165}Dy(NO_3)_3$ with sodium borohydride solution in 0.2N NaOH. Coulter particle analyzer exhibited mean particle size of $2.6{\mu}m$ (range $1{\sim}6{\mu}m$), Even though the $^{165}Dy-MA$ suspension in saline was stored at $37^{\circ}C$ for 24 hours or autoclaved at $121^{\circ}C$ for 30minutes, there was no significant change in particle size and leakage problem indicating the prepared $^{165}Dy-MA$ is sufficiently stable. In-vivo retention studies were carried out by administering $^{165}Dy-MA$ into the knee joint space of normal rabbits. Gamma camera analysis showed high retention in joint space of normal rabbits. Gamma camera analysis showed high retention in joint space even at 24 hours after administration (> 99.9%). The ease with which the $^{165}Dy-MA$ can be made in the narrow size range and their high invitro and vivo stability make them attractive agents for radiation synovectomy.

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Validation of an Enzyme-Immunoassay for Calcitriol in Human Plasma and Evaluation of Its Pharmacokinetics after Single-dose in Korean Volunteers (인체 혈장 중 칼시트리올의 효소면역 분석법 검증 및 단회투여 후 약물동태 연구)

  • Kim, Ye-Tae;Jin, Su-Eon;Kim, Hyun-Ki;Shin, Baek-Ki;Jeong, Ui-Hyeon;Kim, Chong-Kook;Park, Jeong-Sook
    • Journal of Pharmaceutical Investigation
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    • v.39 no.4
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    • pp.309-314
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    • 2009
  • An enzyme immunoassay (EIA) was validated for quantitation of cacitriol in human plasma. Calcitriol was immunoextracted with immunocapsules, which contain monoclonal antibodies to calcitriol linked to solid phase particles in suspension with a vitamin D binding protein inhibitor. Calcitrol was eluated and the eluates were evaporated under a gentle stream of nitrogen gas. The absorbance of analytes was determined using a microplate reader (reference wavelength 650 nm; measurement wavelength 450 nm). The method was specific and sensitive enough to detect as low as 6.5 pmol/L of calcitriol. Linear calibration range was 6.5-491 pmol/L with correlation coefficient greater than 0.99. The overall accuracy was in the range of 83.8 to 111.2% and precision C.V. (%) 0.99 to 8.47%. The recovery was approximately 100% and stability was confirmed during storage and sample preparation. The pharmacokinetic parameters were calculated by baseline subtraction because calcitriol is an endogenous material. Following oral dose of calcitriol, the mean AUC$_{24h}$ was 1038${\pm}$539 pmol/Lhr and C$_{max}$ of 128${\pm}$63.1 pmol/L was reached at 3.50${\pm}$1.07 hr. The mean t$_{1/2}$ of calcitriol was 5.13${\pm}$2.10 hr. The present EIA method was successfully applied to study bioavailability after oral administration of 2 ${\mu}$g of calcitriol in healthy Korean subjects.

Design of Large-scale Drilled Shaft (대구경 현장타설말뚝의 설계 사례)

  • Im, Chul-O;Choi, Young-Seok;Kwak, Ki-Seok;Jang, Hak-Sung
    • Proceedings of the Korean Geotechical Society Conference
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    • 2009.09a
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    • pp.545-553
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    • 2009
  • A lot of long-span marine bridge, which connects land to island or island to island, are being designed and constructed lately in south-west coast in South Korea. In the past, caisson foundations in marine were mainly adopted in construction and stability aspect, however, nowadays with development of pile construction technology, drilled shaft foundations are mainly adopted. As the long span cable stayed bridge and suspension bridge applied with lots of loads are being designed, the scale of pile foundations are getting larger. As the construction cost of substructure including foundation in marine bridges is too high, the appropriate evaluation of the axial bearing capacity of pile becomes a core factor to decide the construction cost of foundation if the drilled shaft is adopted as foundation type of bridge. The evaluation values of skin friction and end bearing capacity of drilled shaft in weathered rock suggested in south Korea are only to introduce the foreign specifications, and most of them are designed in a kind of hard soil layer. Also the allowable load of pile section is less than the expected bearing capacity of pile in the soil condition since the allowable capacity of pile is undervalued. Recently in order to improve this factor the bi-axial hydraulic load test of pile was taken, the data of load transfer analysis of pile, unit of skin friction and end bearing capacity are accumulated. In our country, the design of piles are made with ASD, however, LRFD considering service, strength and extreme state was adopted in Incheon Grand Bridge implemented with BTL, and the research to systematize the resistance coefficient appropriate at home country are being progressed.

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