• Title/Summary/Keyword: purification efficacy

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계란난황항체의 Edwardsiella tarda에 대한 효능 (Efficacy of Egg Yolk Immunoglobulin (IgY) against Edwardsiella tarda Infection)

  • 김영대;오명주;정성주
    • 한국해양바이오학회지
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    • 제1권3호
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    • pp.218-223
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    • 2006
  • 어류의 세균성질병 원인체인 Edwardsiella tarda 감염의 예방과 치료에 E. tarda로 면역한 계란으로부터 얻어진 난황항체 (IgY)의 잠정적인 사용을 평가하였다. PEG법으로 정제된 IgY는 64 kDa의 heavy chain과 27 kDa의 light chain을 가지고 있었다. IgY는 사료 공급 후 위의 pH가 3.4로 낮아지는 2시간째에는 대부분의 활성을 상실하였으나, 장내에서는 IgY의 활성이 나타났다. IgY의 효능 실험 결과 감염 초기에 IgY를 20 mg/fish로 공급한 그룹은 모든 실험에서 대조 그룹에 비해 높은 생존율을 나타내었으며, 장기 내에서 균의 감염 정도 역시 낮았다. 그러나 감염 후기에서는 생존율에 있어서의 유의적인 차이를 확인하기 힘들었다. 이는 넙치의 위에서 대부분의 IgY가 활성을 잃어 낮은 농도만이 장으로 이동하기 때문일 것으로 사료되며, 이 농도에서 IgY는 E. tarda에 대한 뚜렷한 예방이나 치료의 효과 보다는 감염시기를 어느 정도 연장시키는 역할을 하는 것으로 추정된다.

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Evaluation of Recombinant SAG1, SAG2, and SAG3 Antigens for Serodiagnosis of Toxoplasmosis

  • Khanaliha, Khadijeh;Motazedian, Mohammad Hossein;Kazemi, Bahram;Shahriari, Bahador;Bandehpour, Mojgan;Sharifniya, Zarin
    • Parasites, Hosts and Diseases
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    • 제52권2호
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    • pp.137-142
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    • 2014
  • Serologic tests are widely accepted for diagnosing Toxoplasma gondii but purification and standardization of antigen needs to be improved. Recently, surface tachyzoite and bradyzoite antigens have become more attractive for this purpose. In this study, diagnostic usefulness of 3 recombinant antigens (SAG1, SAG2, and SAG3) were evaluated, and their efficacy was compared with the available commercial ELISA. The recombinant plasmids were transformed to JM109 strain of Escherichia coli, and the recombinants were expressed and purified. Recombinant SAG1, SAG2, and SAG3 antigens were evaluated using different groups of sera in an ELISA system, and the results were compared to those of a commercial IgG and IgM ELISA kit. The sensitivity and specificity of recombinant surface antigens for detection of anti-Toxoplasma IgG in comparison with commercially available ELISA were as follows: SAG1 (93.6% and 92.9%), SAG2 (100.0% and 89.4%), and SAG3 (95.4% and 91.2%), respectively. A high degree of agreement (96.9%) was observed between recombinant SAG2 and commercial ELISA in terms of detecting IgG anti-Toxoplasma antibodies. P22 had the best performance in detecting anti-Toxoplasma IgM in comparison with the other 2 recombinant antigens. Recombinant SAG1, SAG2, and SAG3 could all be used for diagnosis of IgG-specific antibodies against T. gondii.

Enhancement of potency and stability of human extracellular superoxide dismutase

  • Kim, Sunghwan;Kim, Hae-Young;Kim, Jung-Ho;Choi, Jung-Hye;Ham, Won-Kook;Jeon, Yoon-Jae;Kang, Hara;Kim, Tae-Yoon
    • BMB Reports
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    • 제48권2호
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    • pp.91-96
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    • 2015
  • Cells express several antioxidant enzymes to scavenge reactive oxygen species (ROS) responsible for oxidative damages and various human diseases. Therefore, antioxidant enzymes are considered biomedicine candidates. Among them, extracellular superoxide dismutase (SOD3) had showed prominent efficacy against asthma and inflammation. Despite its advantages as a biomedicine, the difficulty in obtaining large quantity of active recombinant human SOD3 (rhSOD3) has limited its clinical applications. We found that a significant fraction of over-expressed rhSOD3 was composed of the inactive apo-enzyme and its potency against inflammation depended on the rate of metal incorporation. Also, purified rhSOD3 was unstable and lost its activity very quickly. Here, we suggest an ideal preparative method to express, purify, and store highly active rhSOD3. The enzymatic activity of rhSOD3 was maximized by incorporating metal ions into rhSOD3 after purification. Also, albumin or polyethylene glycol prevented rapid inactivation or degradation of rhSOD3 during preparative procedures and long-term storage.

상수도 급수방식 전환의 타당성 분석 및 최적 범위 산정모델 연구 (A Study on Feasibility Analysis and Optimum Range Calculation Model by Conversion of Water Supply System)

  • 박준열;신휘수;서지원;김기범;구자용
    • 상하수도학회지
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    • 제31권2호
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    • pp.177-186
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    • 2017
  • This study concerned the analysis on the efficiency of the conversion of water tank type supply system to direct water supply system to examine the feasibility of the conversion, as well as the calculation of optimal conversion range that enables the supply of safe, high-quality water at stable pressure in accordance with the standards of water supply facility. The results of this research showed that when converting water supply system from water tank type supply system to direct water supply system, more nodal points could be properly converted and more reduction of electricity usage was expected in case water pressure rather than residence time was fixed. This means that higher efficacy can be obtained by fixing water pressure when converting water supply system. However, since the number of the locations that received on-spot inspection was small and the electricity usage measured was not exclusively by water supply facility, it is difficult to judge that such reduction of electricity usage accurately represents reduced electricity usage by water supply facility alone. therefore, after having secured on-spot information about a larger number of locations in apartment complexes that have converted water supply system, and utilizing information about electricity usage exclusively by water supply facility, the proposed method of this research could be applied to accurately deducing expected reduction of electricity usage by water supply facilities of various other apartment complexes. It is also considered possible to deduce an effective operation method of water supply system by finding out an area that shows low pressure or low residual chlorine concentration in the optimal conversion range of water supply, followed by estimating the proper location of pumping station or the proper chlorine dosage at the power purification plant that supply water to the target area.

시화호 인공습지의 수질정화 및 암모니아 산화균의 분포 연구 (The Efficacy of Water Purification and Distribution of Ammonia Oxidizing Bacteria in Shihwa Constructed Wetland)

  • 김세윤;김미순;이성희;임미영;이영민;김지열;고광표
    • 한국물환경학회지
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    • 제26권1호
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    • pp.10-18
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    • 2010
  • Water quality and the distribution of ammonia oxidizing bacteria were characterized in constructed wetland of Shihwa lake. Both physico-chemical parameters and fecal indicator microorganisms including total coliforms, E.coli, Enterococcus spp. were measured. In addition, denaturant gradient gel electrophoresis (DGGE) was carried out after PCR amplification of amoA gene from input, output, and wetland sites of the Banwol, Donghwa, and Samhwa stream in Shihwa lake area. Physico-chemical parameters were in proper range for typical nitrifying bacteria to grow and perform their biological activities. Average concentrations of fecal indicator microorganisms of wetland samples were lower than those of input sites. These results suggested that microbial water quality improved by the process of constructed wetland. According to phylogenetic information obtained from DGGE from study sites, distribution of nitrifying bacteria from each of input, output, and wetland were generally distinctive one another. In addition, distribution of nitrifying bacteria between Banwol and Donghwa streams showed higher similarity (52.6%) than this of Samhwa stream (15.2%). These results indicated that characteristics of ammonia oxidizing bacteria in Samhwa were unique in comparison with those of Banwol and Donghwa stream.

중소기업융합학회 수록 논문의 연구동향에 대한 계량서지학적 분석 (Bibliometric Analysis on the Research Trends in Journal of Convergence for Information Technology)

  • 김신희
    • 융합정보논문지
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    • 제10권7호
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    • pp.122-130
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    • 2020
  • 본 연구는 중소기업융합학회에 수록된 논문을 대상으로 계량서지학적 분석을 실시함으로써 융합연구에 대한 동향 및 추세를 파악하는 것을 목적으로 한다. 2012~2019년까지 수록된 792편의 논문자료를 수집하였으며, 텍스트 마이닝을 통해 명사 및 복합명사를 추출하고 3차의 전처리 및 정제과정을 거쳐 중심성을 활용한 연결망 분석을 실시하였다. 핵심 연구결과에 따르면 첫째, 2016년 이후 연구의 양적·질적 측면이 향상되었으며, 영향, 융합, 매개효과, 만족, 효과연구가 중점을 이루었다. 둘째, 전반기(2012~2015년)에는 정보, 시스템, 보안 등의 주제를 토대로 공학 및 기술측면의 연구가 집중적으로 수행되어졌다. 셋째, 후반기(2016~2019년)에는 대학생, 부모, 청소년 등을 대상으로 직무, 자기효능감, 교육, 만족, 우울, 스트레스 등을 주제로 연구범위가 확장되어졌다. 본 연구결과는 기존의 연구동향을 파악하는 것과 더불어 신규 연구영역의 확장에 필요한 정보를 제공하는데 의의가 있다.

Cycloinulooligosaccharide fructanotransferase의 결손변이효소에 의한 cyclofructan의 고효율 생산 (High Yield Production of Cyclofructan by Deletion Mutant Enzyme of Cycloinulooligosaccharide Fructanotransferase)

  • 박정하;권현주;김병우
    • 생명과학회지
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    • 제16권1호
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    • pp.1-5
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    • 2006
  • 본 연구는 Penibacillus polymyxa 균주의 CFTase 유전자를 결손 변이시킨 고효율 효소 CFT108을 이용하여 cyclofructan (CF)의 대량생산 조건을 검토하고 생산된 CF의 순수 분리 정제 공정을 개발하였다. CF의 대량생산 조건은 $2\%$의 inulin 기질과 40 unit/g inulin의 기질에서 3시간 반응시켰을 때 최대 생산량 9.5 g/l의 수율을 달성할 수 있었으며, 이때의 in-ulin의 CF 전환율은 $47.5\%$였다. 생산된 CF를 순수 분리 정제하기 위해서 CFTase 반응액 을 exoinulinase 1 unit/ml로 6시간 처리하여 미 반응 inulin을 단당화시켰으며, 유리된 fructose는 $3\%$ CaO로 $CO_2$가스 하에서 $80^{\circ}C$, 10분간 3회 흡착제거 시킴으로써 순수 정제하였다. 정제된 CF를 HPLC로 확인한 결과 정제도는 $95\%$ 이상이였으며, $CF_6,\;CF_7,\;CF_8$ 비율이 72 : 27 : 1 이였다.

An Outer Membrane Protein Preparation as a Vaccine against Pseudomonas aeruginosa Infection

  • Park, Wan-Je;Cho, Yang-Je;Ahn, Dong-Ho;Jung, Sang-Bo;Lee, Na-Gyong;Kim, Hyun-Su;Hahm, Kyung-Soo;Kim, Yu-Sam
    • Journal of Microbiology and Biotechnology
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    • 제7권2호
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    • pp.144-150
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    • 1997
  • We developed a simple and efficient method to prepare a Pseudomonas vaccine of outer membrane (OM) proteins free from lipopolysaccharide (LPS). A three step purification process including extraction, ultrafiltration and ultracentrifugation effectively removed LPS from the OM protein fraction. Approximately 2 mg of the OM proteins was obtained from 1 g of wet cell. LPS contaminant in the vaccine preparation was less than 0.003% (w/w) of protein and protease activity was not detectable. To achieve a wide range of protection, OM proteins prepared from four attenuated P. aeruginosa strains were mixed in equal amounts and used as a vaccine, which elicited in rabbits a high titer of antibody reactive to all of the seven Fisher types. The antisera from the immunized rabbit had a strong reactivity to vaccine proteins larger than 25 kDa. In a burned mouse infection model, immunization with the vaccine significantly enhanced bacterial clearance in the Pseudomonas infected skin. The vaccination also provided mice an excellent protection against Pseudomonas infection (11, 16). Data on antigenicity, mutagenicity, acute, subacute toxicity and pharmacological tests confirmed the safety of the vaccine (1, 3, 10, 12, 17). These data demonstrate that this method can be applied to manufacture a bacterial vaccine of OM proteins with safety and prophylactic efficacy at a practical low cost.

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Human growth hormone의 개발과 이에 따른 효능 및 안전성 평가 (Development of Recombinant Human Growth Hormone in Yeast: Efficacy Evaluation and Safety Assessment)

  • 이상균
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 1997년도 국제심포지움 및 추계학술대회
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    • pp.38-42
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    • 1997
  • 인간성장 호르몬은 191개의 아미노산으로 구성된 펩타이드 호르몬의 일종으로 사람의 뇌하수체에서 분비된다. 본연구의 목적은 저렴한 가격의 재조합 성장호르몬을 안정적으로 공급함은 물론 부작용을 최대한으로 줄이는데 있다. 효모에서의 성장호르몬 재조합 과정은 유전자조작과 이에따른 정제기술의 확립을 시작으로 물리 화학적 또는 생물학적 물성 연구, 다양한 전임상 시험 및 임상 시험의 단계를 수행한 후, 인허가 과정을 거쳐서 완료되었다. 다양한 실험 결과에 의하면 당소에서 재조합 기술로 생산한 고순도의 인간성장 호르몬은 안정된 물성을 보유하고 있을 뿐만아니라 전임상 시험 결과 인체 투여용량의 수십배의 고용량을 투여하여도 별다른 독성 증상을 관찰할 수 없었으며, 임상 시험에서도 대상 환아의 신장이 지속적으로 성장하는 등 만족할 만한 결과를 보여 주었다.

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Minor Coat Protein pIII Domain (N1N2) of Bacteriophage CTXф Confers a Novel Surface Plasmon Resonance Biosensor for Rapid Detection of Vibrio cholerae

  • Shin, Hae Ja;Hyeon, Seok Hywan;Cho, Jae Ho;Lim, Woon Ki
    • 한국미생물·생명공학회지
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    • 제49권4호
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    • pp.510-518
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    • 2021
  • Bacteriophages are considered excellent sensing elements for platforms detecting bacteria. However, their lytic cycle has restricted their efficacy. Here, we used the minor coat protein pIII domain (N1N2) of phage CTXφ to construct a novel surface plasmon resonance (SPR) biosensor that could detect Vibrio cholerae. N1N2 harboring the domains required for phage adsorption and entry was obtained from Escherichia coli using recombinant protein expression and purification. SDS-PAGE revealed an approximate size of 30 kDa for N1N2. Dot blot and transmission electron microscopy analyses revealed that the protein bound to the host V. cholerae but not to non-host E. coli K-12 cells. Next, we used amine-coupling to develop a novel recombinant N1N2 (rN1N2)-functionalized SPR biosensor by immobilizing rN1N2 proteins on gold substrates and using SPR to monitor the binding kinetics of the proteins with target bacteria. We observed rapid detection of V. cholerae in the range of approximately 103 to 109 CFU/ml but not of E. coli at any tested concentration, thereby confirming that the biosensor exhibited differential recognition and binding. The results indicate that the novel biosensor can rapidly monitor a target pathogenic microorganism in the environment and is very useful for monitoring food safety and facilitating early disease prevention.