• Title/Summary/Keyword: Fab'

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Effects of Disulfide and Thioether Linkages on Stability and Cytotoxicity of Anti-CALLA Fab-Ricin A Immunotoxins

  • Lee, Jung-Tae;Woo, Byung-Ho;Lee, Kang-Choon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.277-277
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    • 1996
  • Anti-CALLA Fab-RTA immunotoxins were constructed using two crosslinking agent, SMPT and SMCC, to generate a disulfide and a thioether bridge between Fab fragment of K269-65 MoAb and RTA toxin moieties, respectively. These immunotoxins were selectively immunoreative with CALLA$\^$+/ B-lineage Daudi cells. SMPT and SMCC mediated RTA immunotoxins were prepared with 49% and 53% of the RTA conjugation yields, respectively. SDS-PAGE analysis show that immunotoxins were constructed with major Fab-1 RTA (76kda), minor Fab-2RTA (106kda) and Fab-3RTA (136kda) compositions. The breakdown rates of immunotoxins were determined in the presence of glutathione by measuring the amount of reduced immunotoxins using size-exclusion HPLC. The SMCC immunotoxins were more resistant to the glutathione than SMPT immunotoxins. But, our data showed that the SMPT mediated disulfide bonded immunotoxins were much more active than the SMCC mediated thioether bonded immunotoxins to kill the target cells in vitro.

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Review for Retrospective Exposure Assessment Methods Used in Epidemiologic Cancer Risk Studies of Semiconductor Workers: Limitations and Recommendations

  • Park, Donguk
    • Safety and Health at Work
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    • v.9 no.3
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    • pp.249-256
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    • 2018
  • This article aims to provide a systematic review of the exposure assessment methods used to assign wafer fabrication (fab) workers in epidemiologic cohort studies of mortality from all causes and various cancers. Epidemiologic and exposure-assessment studies of silicon wafer fab operations in the semiconductor industry were collected through an extensive literature review of articles reported until 2017. The studies found various outcomes possibly linked to fab operations, but a clear association with the chemicals in the process was not found, possibly because of exposure assessment methodology. No study used a tiered assessment approach to identify similar exposure groups that incorporated manufacturing era, facility, fab environment, operation, job and level of exposure to individual hazardous agents. Further epidemiologic studies of fab workers are warranted with more refined exposure assessment methods incorporating both operation and job title and hazardous agents to examine the associations with cancer risk or mortality.

Stability and Cytotoxicity of Fab-Ricin A Immunotoxins Prepared with Water Soluble Long Chain Heterobifunctional Crosslinking Agents

  • Woo, Byung-Ho;Lee, Jung-Tae;Park, Myung-Ok;Lee, Kang-Ro;Han, Jeung-Whan;Park, Eun-Seok;Yoo, Sun-Dong;Lee, Kang-Choon
    • Archives of Pharmacal Research
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    • v.22 no.5
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    • pp.459-463
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    • 1999
  • The effects of the hindered and non-hindered water soluble long-chain disulfide bonds on the stability and cytotoxicity of the ricin A chain (RTA) immunotoxin were examined. The RTA immunotoxins were prepared with the Fab fragments of anti-common acute lymphoblastic leukemia antigen (CALLA) monoclonal antibody (Fab-RTA) using sulfosuccinimidyl-6-[(-methyl-(-2-pyridyldithio)toluamido]toluamido]hexanoate (S-LC-SMPT) and sulfosuccinimidyl-6-[3-(2-pyridyldithio-propionamido]hexanoate (S-LC-SPDP). The prepared Fab-RTA immunotoxins were evaluated for their conjugation yield, immunoreactivity, thermal and disulfide bond stability and cytotoxicity. The conjugation yield of the Fab-RTA immunotoxin from the water soluble long chain crosslinking agents, S-LC-SMPT and S-LC-SPDP, were comparable. Both Fab-RTA immunotoxins exhibited a similar immunoreactivity and thermal stability in aqueous solution. However, S-LC-SMPT -mediated Fab-RTA, sterically hindered, showed an enhanced disulfide bond stability in vitro over S-LC-SPDP mediated one. In the cytotoxicity against antigenic cell Daudi, the S-LC-SMPT -mediated RTA immunotoxin maintained a comparable cytotoxicity, compared with S-LC-SPDP mediated Fab-RTA immunotoxin.

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A Divalent Immunotoxin Formed by the Disulfide Bond between Hinge Regions of Fab Domain

  • Choe, Seong Hyeok;Kim, Ji Eun;Lee, Yong Chan;Jang, Yeong Ju;Choe, Mu Hyeon
    • Bulletin of the Korean Chemical Society
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    • v.22 no.12
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    • pp.1361-1365
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    • 2001
  • Recombinant immunotoxins are hybrid cytotoxic proteins designed to selectively kill cancer cells. A divalent immunotoxins, [B3(FabH1)-PE38]2, was constructed by recombining Fab domain of B3 antibody as a cell-targeting domain and Pseudo monas exotoxin A (PE) as a cytotoxic domain. Monoclonal antibody, B3, is the murine antibody (IgG1k) directed against Lewis Y-related carbohydrate antigens, which are abundant on the surface of many carcinomas. Fab fragment of this antibody was used in this study with the modified hinge sequence where last two cysteines out of three were mutated to serine. PE is a 66 kDa bacterial toxin that kills eukaryotic cells by inhibiting protein synthesis with ADP ribosylation of ribosomal elongation factor 2 (EF2). Fc region of B3 antibody was substituted with the truncated form of PE (38 kDa, PE38) on DNA level. [B3(FabH1)-PE38]2 was formed by disulfide bond between cysteines in the modified hinge region of B3(FabH1)-PE38. Each polypeptide for recombinant immunotoxins was overexpressed in Escherichia coli and collected as inclusion bodies. Each inclusion body was solubilized and refolded, and cytotoxic effects were measured. Divalent immunotoxins, [B3(FabH1)-PE38]2, had ID50 values of about 10 ng/mL on A431 cell lines and about 4 ng/mL on CRL1739 cell lines. Control immunotoxins, B3(scFv)-PE40, had ID50 values of about 28 ng/mL on A431 cell lines and about 41 ng/mL on CRL1739 cell lines. Divalent immunotoxins, [B3(FabH1)-PE38]2, had higher cytotoxic effects than B3(scFv)-PE40 control immunotoxins.

Polymer Wafer bonding of MEMS device and Cap Wafer with deep cavity (Deep cavity를 가진 Cap Wafer와 MEMS 소자의 Polymer Wafer bonding)

  • Lee, Hyun-Kee;Park, Tae-Joon;Yoon, Sang-Kee;Park, Nam-Su;Park, Hyung-Jae;Min, Jong-Hwan;Lee, Yeong-Gyu
    • Proceedings of the KIEE Conference
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    • 2011.07a
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    • pp.1702-1703
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    • 2011
  • MEMS 소자의 Wafer level Package 관련하여 Deep cavity를 가진 Cap Wafer와 Polymer bonding 중 cavity 단차로 인한 Polymer Patterning 및 접합 불량의 어려움을 극복할 수 있는 새로운 공정 flow를 제안하였다. Cavity를 형성할 때 사용하는 Si deep etching Mask인 기존의 Photoresist를 접합용 감광성 Polymer로 대체하고, cavity 형성 후, 별도의 추가 공정 없이 이 Polymer를 이용해 Wafer bonding을 진행하였다. 이를 통해 cavity 단차에 따른 문제를 해결함과 동시에 공정이 단순하고 제작 비용이 저렴하며, 신뢰성 있는 Wafer level Package를 구현하였다.

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Preparation and Characteristics of Phosphatidylcholine Liposomes Covalently Coupled with Immunoglobulin Fragments (이뮤노글로부린 토막과 결합된 포스파티딜콜린 리포솜의 제조와 특성)

  • Park, Jung-Hwan;Lee, Eun-Ok;Kim, Jong-Duk
    • Journal of Pharmaceutical Investigation
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    • v.22 no.2
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    • pp.115-124
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    • 1992
  • The characteristics and stabilities of phosphatidylcholine liposomes covalently coupled with immunoglobulin fragments prepared by the REV method were investigated by the dynamic light scattering, absorbance and calcein release. Using a sulfhydryl-reactive phospholipid derivative of N-[4$({\rho}-maleimido-phenyl)$ butyl] phosphatidylethanolamine (MPB-PE), Fab' antibody fragments were covalently combined with preformed large unilamellar vesicles (LUV), Coupling ratio was $250\;{\mu}g$ of $Fab'/{\mu}mol$ of phospholipid in vesicles, From dynamic light scattering, it was found that the size of the vesicles increases as the ratio of cholesterol to lipid increases, but that apparently, the size of liposomes was not sensitive to the existence of Fab' fragments. Regardless of inserting Fab' fragments, the absorbance of liposomes decreased as the amounts of bile salt (BS) added. At very low BS concentrations, BS/lipid aggregates would be formed in the outer vesicles monolayer, while, at the high BS concentrations, mixed micelles would be preferred. The vesicles incorporated with Fab' fragments, however, are more resistant to the bile salts than the MPB-PE vesicle are. The absorbance of vacant liposomes and calcein release resulted in that the Fab' vesicles and MPB-PE vesicles by the REV method are very stable, but that those by the sonication method sufferred the significant change of turbidities.

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The Effects of Growth Temperature and Substrate Tilt Angle on GalnP/GaAs Tandem Solar Cells

  • Jun, Dong-Hwan;Kim, Chang-Zoo;Kim, Hog-Young;Shin, Hyun-Beom;Kang, Ho-Kwan;Park, Won-Kyu;Shin, Ki-Soo;Ko, Chul-Gi
    • JSTS:Journal of Semiconductor Technology and Science
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    • v.9 no.2
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    • pp.91-97
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    • 2009
  • The performance of GaInP/GaAs tandem solar cells with AlInP growth temperatures of 680$^{\circ}C$ and 700 $^{\circ}C$ on n-type GaAs (100) substrate with 2$^{\circ}$ and 6$^{\circ}$ tilt angles has been investigated. The series resistance and open circuit voltage of the fabricated tandem solar cells are affected by the substrate tilt angles and the growth temperatures of the window layer when zinc is doped in the tunnel diode. With carbon doping as a p-type doping source in the tunnel diode and the effort of current matching between top and bottom cells, GaInP/GaAs tandem solar cell has been exhibited 25.58% efficiency.

Methyl-Branched Fatty Acids, Inhibitors of Enoyl-ACP Reductase with Antibacterial Activity from Streptomyces sp. A251

  • Zheng, Chang-Ji;Sohn, Mi-Jin;Chi, Seung-Wook;Kim, Won-Gon
    • Journal of Microbiology and Biotechnology
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    • v.20 no.5
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    • pp.875-880
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    • 2010
  • Bacterial enoyl-ACP reductase (FabI) has been demonstrated to be a novel antibacterial target. In the course of our screening for FabI inhibitors, we isolated two methyl-branched fatty acids from Streptomyces sp. A251. They were identified as 14-methyl-9(Z)-pentadecenoic acid and 15-methyl-9(Z)-hexadecenoic acid by MS and NMR spectral data. These compounds inhibited Staphylococcus aureus FabI with $IC_{50}$ values of 16.0 and $16.3\;{\mu}M$, respectively, but did not affect FabK, an enoyl-ACP reductase of Streptococcus pneumonia, at $100\;{\mu}M$. Consistent with their selective inhibition for FabI, they blocked intracellular fatty acid synthesis as well as the growth of S. aureus, but did not inhibit the growth of S. pneumonia. Additionally, these compounds showed reduced antibacterial activity against fabI-overexpressing S. aureus, compared with the wild-type strain. These results demonstrate that the methylbranched fatty acids show antibacterial activity by inhibiting FabI in vivo.

AR system for FAB construction management using BIM data under fast track condition (패스트트랙 환경에서 FAB신축을 지원하는 BIM기반 AR 시스템 개발)

  • Lee, Sang-Won;Lee, Kwang-Soo;Choi, Sung-In;Ryu, Seong-Chan;Park, Jung-Seo
    • Journal of KIBIM
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    • v.12 no.4
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    • pp.1-18
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    • 2022
  • New Fabrication Facility (FAB) construction is performed with Building Information Modeling (BIM) based design. The BIM design data keep updated during the FAB construction. To improve fast-track construction management, a Fabrication Facility Augmented Reality (FABAR) was developed. This study introduces a FABAR system development process and shows performance evaluation results of the FABAR prototype system. The FABAR is implemented with three major modules: Augmented Reality (AR) visualization unit (Room-box) to transfer big BIM data to AR data, AR registration and tracking unit to match AR with real scape and to keep AR coordination in real, and AR data management unit to enhance usability. The prototype performance results were as follows: visualization of design BIM data via AR within 24 hours, precise AR registration and tracking registration, and appropriate usability to support FAB construction management at site. The results indicate that the FABAR is applicable for FAB construction management. Especially, the BIM data transformation method using Room-box in this study signifies a new construction management approach using fluctuating BIM design data in the fast track construction condition.

An Analytical Method of Thromboxane $B_2$ by Fast Atom Bombardment Mass Spectrometry (고속원자충격질량분석법을 이용한 Thromboxane $B_2$ 분석)

  • Jang, Suk-Yoon;Kim, Jung-Hoon;Lee, Yong-Moon;Jang, Seung-Ki;Moon, Dong-Cheul
    • Analytical Science and Technology
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    • v.6 no.4
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    • pp.349-357
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    • 1993
  • Analytical methods of thromboxane $B_2(TXB_2)$ using various techniques of Fast Atom Bombardment mass spectrometry (FAB MS) were studied, static FAB condition was investigated to obtain linear response curve using docosanoic acid as a internal standard. For maximum sensitivity, a continuos-flow(CF-) FAB MS by selected-ion monitoring(SIM) with devised sample introduction system, has been developed to quantiate thromboxane $B_2$ in biological sample. Instrumental parameters affecting sensitivity, reproducibility has been studied. The method has been optimized with respect to the eluent, 0.75% glycerol(in EtOH v/v) and flow rate of $3.7{\mu}l/min.$ Under the condition, detection limits were below 10pg in SIM mode and a good linear relationship between dose and response was achieved.

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