• 제목/요약/키워드: fab E

검색결과 53건 처리시간 0.034초

Bacillus subtilis subsp. krictiensis가 생산하는 항진균 물질 KRF-001의 구조 결정 (Structure Determination of Antifungal KRF-001 Produced by Bacillus subtilis subsp. krictiensis)

  • 김성기;이남경;정태숙;김영국;최진자;복성해
    • 한국미생물·생명공학회지
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    • 제19권6호
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    • pp.598-603
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    • 1991
  • 신규 미생물 Bacillus subtilis subsp. krictiensis의 배양액으로부터 식물 및 인체 병원균에 폭넓은 항진균 활성을 나타내는 DRF-001으로 총칭되는 6개의 cyclic peptide의 복합체(A에서 F)를 분리하였다. 이 6개 peptides의 분자량은 FAB-MS 측정결과, A가 1042, B와 C가 1056, D와 E가 1070 그리고 F가 1084였다. 이 복합체의 구조를 각종 자기분석을 통해 해석한 결과, 구조 중에 1몰씩의 glutamine, proline, tyrosine, serine 및 unusal $\beta$-amino acid와 3몰의 asparagine을 공통적으로 갖고 있으나, $\beta$-amino acid의 탄소수와 말단 methyl 구조에 차이점이 있음을 알 수있었다. 구성 amino acid들의 서열과 입체구조 결정에 의하여 KRF-001의 잠정구조를 결정하였다.

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반도체 Fab의 생산운영시스템 구축을 위한 상황적응형 디스패칭 방법론 : 공정전환시간이 있는 장비를 중심으로 (An Adaptive Dispatching Architecture for Constructing a Factory Operating System of Semiconductor Fabrication : Focused on Machines with Setup Times)

  • 정근채
    • 산업공학
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    • 제22권1호
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    • pp.73-84
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    • 2009
  • In this paper, we propose a dispatching algorithm for constructing a Factory Operating System (FOS) which can operate semiconductor fabrication factories more efficiently and effectively. We first define ten dispatching criteria and propose two methods to apply the defined dispatching criteria sequentially and simultaneously (i.e. fixed dispatching architecture). However the fixed type methods cannot apply the criteria adaptively by considering changes in the semiconductor fabrication factories. To overcome this type of weakness, an adaptive dispatching architecture is proposed for applying the dispatching criteria dynamically based on the factory status. The status can be determined by combining evaluation results from the following three status criteria; target movement, workload balance, and utilization rate. Results from the shop floor in past few periods showed that the proposed methodology gives a good performance with respect to the productivity, workload balance, and machine utilization. We can expect that the proposed adaptive dispatching architecture will be used as a useful tool for operating semiconductor fabrication factories more efficiently and effectively.

Characterization of Extremely Hydrophobic Immunostimulatory Lipoidal Peptides by Matrix Assisted Laser Desorption Ionization Mass Spectrometry

  • 장정석;이성택;장윤석
    • Bulletin of the Korean Chemical Society
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    • 제17권11호
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    • pp.1036-1039
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    • 1996
  • Synthetic lipoidal peptides based on viral protein sequences have been prepared. These peptides contain an N-palmitoyl group at the N-terminal residue, which is a modified cysteine, containing a S-[2,3-bis(acyloxy)-(2-R,S)-propyl] moiety. When this residue (Pam3Cys) is at the N-terminus of a synthetic peptide, it acts as potent immunoadjuvant to enhance both IgM and IgG antibody responses to the attached peptide. Conventional analytical procedures (e.g., Edman degradation and amino acid analysis) are either not applicable due to the N-terminal modification, or do not provide confirmation of the intact structure. Chromatographic analysis is also hindered by the tendency of these lipoidal Pam3Cys peptides to form large aggregates, and in some cases to be permanently adsorbed on reversed phase columns. We have applied several mass spectrometric techniques, including fast atom bombardment (FAB), electrospray ionization (ESI) and matrix assisted laser desorption ionization (MALDI) to characterize the intact structures of a number of different Pam3Cys synthetic peptides. The MALDI-MS has been found to be the most sensitive for the analysis of the structure of Pam3Cys peptides.

대장균의 회분식 발효에 의해 생산된 덱스트란 결합 파아지를 활용한 설탕 제조공정 오염 검출 (Detection of Sugar Process Contamination Using Dextran Binding hages Produced by Batch Fermentation of Escherichia coli)

  • 김두운
    • 한국식품저장유통학회지
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    • 제15권5호
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    • pp.617-621
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    • 2008
  • Sequential passes through $Sephadex^{TM}$ columns were used to select phages that displays ligands for dextran ($\alpha$-1,6 linked linear chains) from a phage antibody library. Those phages that bound to the $Sephadex^{TM}$ in each iteration were replicated in E. coli. A phage preparation isolated on the third round selection produced 5.4 nephelos turbidity units (NTU) in a dextran specific immunonephelometric assay, a 2.2 fold higher value than the phage preparation from the first round selection. This phage gave $72\;{\pm}\;10$ normalized intensity (N.I.) in a dip-stick assay against high molecular size dextran (T2000, $2\;{\times}\;10^6) and significantly lower color ($30\;{\pm}\;6$ N.I.) against low molecular size dextran (T10, $10^4$). The presence of an Fab insert in each of these phages was confirmed using a $\beta-galactosidase linked assay and polymerase chain reaction.

Three New Flavonoids Of Spatholobus suberectus

  • Yoon, Jeong-Seon;Lim, Song-Won;Yang , Hye-Kyeong;Ahn , Tae-Kyeong;Kim, Eun-Sil;Sung, Sang-Hyun;Kim, Young-Choong
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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    • pp.202.1-202.1
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    • 2003
  • Six compounds were isolated from the 90% MeOH fraction of the vine stem of Spatholobus suberectus Dunn (Leguminosae) using silica gel, reverse phase column chromatography and RP-HPLC. Structures of compounds 1-6 were elucidated by spectroscopic parameters of IR, E1 MS, FAB MS, lD-NMR and 2D-NMR spectrum and identified as pseudobaptigenin (1), genistein (2), (2R)-7-hydroxy-6-methoxyflavanone (3), (3R,4R)-2",4"-dihydroxy-6,7-methylenedioxy-isoflavan-4-ol (4), (3R,4R)-7,2" -dihydroxy- 4"-methoxyisoflavan-4-ol (5), sativan (6), respectively. Compounds 3, 4 and 5 have been newly reported in nature.

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Generation and Characterization of Monoclonal Antibodies to the Ogawa Lipopolysaccharide of Vibrio cholerae O1 from Phage-Displayed Human Synthetic Fab Library

  • Kim, Dain;Hong, Jisu;Choi, Yoonjoo;Han, Jemin;Kim, Sangkyu;Jo, Gyunghee;Yoon, Jun-Yeol;Chae, Heesu;Yoon, Hyeseon;Lee, Chankyu;Hong, Hyo Jeong
    • Journal of Microbiology and Biotechnology
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    • 제30권11호
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    • pp.1760-1768
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    • 2020
  • Vibrio cholerae, cause of the life-threatening diarrheal disease cholera, can be divided into different serogroups based on the structure of its lipopolysaccharide (LPS), which consists of lipid-A, core-polysaccharide and O-antigen polysaccharide (O-PS). The O1 serogroup, the predominant cause of cholera, includes two major serotypes, Inaba and Ogawa. These serotypes are differentiated by the presence of a single 2-O-methyl group in the upstream terminal perosamine of the Ogawa O-PS, which is absent in the Inaba O-PS. To ensure the consistent quality and efficacy of the current cholera vaccines, accurate measurement and characterization of each of these two serotypes is highly important. In this study, we efficiently screened a phage-displayed human synthetic Fab library by bio-panning against Ogawa LPS and finally selected three unique mAbs (D9, E11, and F7) that specifically react with Ogawa LPS. The mAbs bound to Vibrio cholerae vaccine in a dose-dependent fashion. Sequence and structure analyses of antibody paratopes suggest that IgG D9 might have the same fine specificity as that of the murine mAbs, which were shown to bind to the upstream terminal perosamine of Ogawa O-PS, whereas IgGs F7 and E11 showed some different characteristics in the paratopes. To our knowledge, this study is the first to demonstrate the generation of Ogawa-specific mAbs using phage display technology. The mAbs will be useful for identification and quantification of Ogawa LPS in multivalent V. cholerae vaccines.

Ti3Ci2Tix MXene 기반 전극 소재의 자가 치유 적용 기술 개발 동향 (Recent Progress of Ti3Ci2Tix MXene Electrode Based Self-Healing Application)

  • 최준상;정승부;김종웅
    • 마이크로전자및패키징학회지
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    • 제30권3호
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    • pp.20-34
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    • 2023
  • 수 나노미터의 두께의 단일 또는 여러 층으로 구성된 2차원 소재는 전기전도성, 유연성, 광학적 투명성 등의 고유한 특성으로 많은 연구 분야에서 활용되고 있다. 이 중 Electronic skin (E-Skin)이나 Smart Textile 과 같은 반복적인 기계적 동작이 수반될 수 있다. 또한, 온도, 습도, 압력과 같은 외부적 요인에 노출이 되는 경우가 빈번하다. 이 때, 소자의 내구성과 수명 저하를 유발하기 때문에 자가 치유 특성이 내포된 소자를 제작하기 위한 연구가 많이 이루어지고 있다. 최근 다양한 2차원 소재 중 자가 치유 기능을 구현할 수 있는 Ti3Ci2Tix MXene 기반 전극의 복합 소재의 연구 결과가 학계의 주목을 받고 있다. 본 논문에서는 Ti3Ci2Tix MXene의 다양한 합성 방법 및 특성에 대해 소개한 후, Ti3Ci2Tix MXene 전극 기반의 자가 치유 적용 기술 사례에 대해 알아보고자 한다.

Expression of a Functional zipFv Antibody Fragment and Its Fusions with Alkaline Phosphatase in the Cytoplasm of an Escherichia coli

  • Hur, Byung-Ung;Choi, Hyo-Jung;Yoon, Jae-Bong;Cha, Sang-Hoon
    • IMMUNE NETWORK
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    • 제10권2호
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    • pp.35-45
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    • 2010
  • Background: Expression of recombinant antibodies and their derivatives fused with other functional molecules such as alkaline phosphatase in Escherichia coli is important in the development of molecular diagnostic reagents for biomedical research. Methods: We investigated the possibility of applying a well-known Fos-Jun zipper to dimerize $V_H$ and $V_L$ fragments originated from the Fab clone (SP 112) that recognizes pyruvate dehydrogenase complex-E2 (PDC-E2), and demonstrated that the functional zipFv-112 and its alkaline phosphatase fusion molecules (zipFv-AP) can be produced in the cytoplasm of Origami(DE3) trxB gor mutant E. coli strain. Results: The zipFv-AP fusion molecules exhibited higher antigen-binding signals than the zipFv up to a 10-fold under the same experimental conditions. However, conformation of the zipFv-AP seemed to be influenced by the location of an AP domain at the C-terminus of $V_H$ or $V_L$ domain [zipFv-112(H-AP) or zipFv-112(L-AP)], and inclusion of an AraC DNA binding domain at the C-terminus of VH of the zipFv-112(L-AP), termed zipFv-112(H-AD/L-AP), was also beneficial. Cytoplasmic co-expression of disulfide-binding isomerase C (DsbC) helped proper folding of the zipFv-112(H-AD/L-AP) but not significantly. Conclusion: We believe that our zipFv constructs may serve as an excellent antibody format bi-functional antibody fragments that can be produced stably in the cytoplasm of E. coli.

고농도 재조합 대장균의 Fed-batch 배양 시스템을 이용한 Pyruvate Dehydrogenase Complex-E2 특이성 인간 모노클론 항체의 생산 (Production of Pyruvate Dehydrogenase Complex-E2 Specific Human Monoclonal Antibody in Fed-batch Culture Systems with High Cell Density Recombinant Escherichia coli)

  • 이미숙;전주미;차상훈;정연호
    • KSBB Journal
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    • 제15권5호
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    • pp.489-496
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    • 2000
  • 고농도 유전자 재조합 대장균을 이용하여 pyruvate dehy-drogenase complex-E2 특이성 인간 모노클론 항체의 Fab 부분을 효율적으로 생산하기 위해 회분식, 이단 연속식, 반 유 가식, two-stage cyclic fed-batch 동 여러 가지 배양 방법이 조사되었다. 먼저 플라스미드 안정성 문제를 극복하기 위해 growth stage와 production stage를 분리하는 two-phase 회분식 배양과 이단 연속식 시스템을 시도하였다 그 결과 two-phase 회분식 배양보다는 이단 연속식 배양에서의 세포농도와 항체 생산성이 우수하였다 또한 이단 연속식 배양에서의 세포 성 장과 항처l 생산성은 용존산소를 제어한 경우가 그렇지 않은 경우보다 월등하게 높았다. 그리고 plasmid 안정성에 있어서 는 실험기간 내에 거의 100%를 유지하여 높은 안정도를 보 여주었다. 유가식 공정에 적합한 공급 배지로 변형된 M9 배 지가 최적배지로 선정되었고 이 배지 중 최적의 CjN 비율을 조사한 결파 2:3으로 결정되었다. 반 유가식 시스템에서 constant feeding 전략을 사용할 경우 최적 공급속도는 $0.6g/\ell/hr$이었다. 또한 pulse에 의해 공급배지를 공급할 경우에는 총 공급 량이 같을 경우 소량으로 자주 공급해 주는 것이 공급배지를 한꺼번에 많은 양을 공급해주는 것 보다 바람직하였다. 여러 가지 feeding 전략을 조사해 본 결과 linear feeding 방법이 가장 효과적이었다. 하지만 linear feeding 방법마저도 고농도 세포배양에 한계가 있었기 때문에 pH-stat 방법을 이용한 two-stage cyclic fed-batch 시스템을 시도하여 $54 g/\ell$의 세포 를 얻을 수 있었다. 따라서 이 방법이 일단 생산성 향상을 위한 세포의 고농도 배양에는 조사한 여러 배양 시스템 중에 가장 효율적인 시스템임올 알 수 있었다 하지만 이 시스템 에서 포도당을 낮은 level로 유지할 수 있었으나, 초산의 과도한 축적으로 항체 생산성의 향상은 예상에 비해 크지 않았다.

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Toxicity Monitoring of Endocrine Disrupting Chemicals (EDCs) Using Freeze-dried Recombinant Bioluminescent Bacteria

  • Kim, Sung-Woo;Park, Sue-Hyung;Jiho Min;Gu, Man-Bock
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제5권6호
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    • pp.395-399
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    • 2000
  • Five different freeze-dried recombinant bioluminescent bacteria were used for the detection of cellular stresses caused by endocrine disrupting chemicals. These strains were DPD2794 (recA::luxCDABE), which is sensitive to DNA damage, DPD2540 (fabA::luxCDABE), sensitive to cellular membrane damage, DPD2511 (katG::luxCDABE), sensitive to oxidative damage, and TV1061 (grpE::luxCDABE), sensitive to protein damage. GC2, which emits bioluminescence constitutively, was also used in this study. The toxicity of several chemicals was measured using GC2. Damage caused by known endocrine disrupting chemicals, such as nonyl phenol, bisphenol A, and styrene, was detected and classified according to toxicity mode, while others, such as phathalate and DDT, were not detected with the bacteria. These results suggest that endocrine disrupting chemicals are toxic in bacteria, and do not act via an estrogenic effect, and that toxicity monitoring and classification of some endocrine disrupting chemicals may be possible in the field using these freeze-dried recombinant bioluminescent bacteria.

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