• 제목/요약/키워드: Efficient Purification

검색결과 214건 처리시간 0.038초

Expression and Efficient One-Step Chromatographic Purification of a Soluble Antagonist for Human Leukemia Inhibitory Factor Receptor in Escherichia coli

  • Kim, Eun-Yeong;Choi, Hee-Jung;Chung, Tae-Wook;Jang, Se Bok;Kim, Kibong;Ha, Ki-Tae
    • Journal of Microbiology and Biotechnology
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    • 제25권8호
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    • pp.1307-1314
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    • 2015
  • Leukemia inhibitory factor (LIF) is a member of the IL-6 cytokine family, having pleiotropic actions such as maintaining stem cell pluripotency and enabling blastocyst implantation. Because the action of LIF is mediated by a ligand-receptor interaction with the LIF receptor (LIF-R), an antagonist for LIF-R has been developed to inhibit LIF-induced signaling. In this study, we present a novel method for the production and purification of an antagonist to human LIF-R (hLA). His-tagged hLA was expressed in E. coli, and simple purification methods without any endopeptidase cleavage were designed. In addition, we determined the optimal temperature conditions for enhancing the production of soluble hLA. Finally, the bioactivity of His-tagged hLA was examined using STAT3 phosphorylation and receptivity of human endometrial ECC-1 cells. Our strategy provides a rapid and efficient method to produce biologically active recombinant hLA.

Development of a Simple Cell Lysis Method for Recombinant DNA Using Bacteriophage Lambda Lysis Genes

  • Jang, Bo-Yun;Jung, Yun-A;Lim, Dong-Bin
    • Journal of Microbiology
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    • 제45권6호
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    • pp.593-596
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    • 2007
  • In this study, we describe the development of a simple and efficient method for cell lysis via the insertion of a bacteriophage lambda lysis gene cluster into the pET22b expression vector in the following order; the T7 promoter, a gene for a target protein intended for production, Sam7 and R. This insertion of R and Sam7 into pET22b exerted no detrimental effects on cellular growth or the production of a target protein. The induction of the T7 promoter did not in itself result in the autolysis of cells in culture but the harvested cells were readily broken by freezing and thawing. We compared the efficiency of the cell lysis technique by freezing and thawing to that observed with sonication, and determined that both methods completely disintegrated the cells and released proteins into the solution. With our modification of pET22b, the lysis of cells became quite simple, efficient, and reliable. This strategy may prove useful for a broad variety of applications, particularly in experiments requiring extensive cell breakage, including library screening and culture condition exploration, in addition to protein purification.

수질 정화 기능 극대화 인공식물섬 개발을 위한 특허 동향 분석 (Analysis of Patents Artificial Floating Island for Maximizing the Development of Water Purification)

  • 김정호;윤용한
    • 한국환경과학회지
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    • 제21권7호
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    • pp.825-835
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    • 2012
  • This study for the development of water purification Artificial floating island maximizing domestic Artificial floating island patent trends and product development, according to the timing of patent registration was analyzed for trends. In addition, domestic invention patent technology Artificial floating island typed according to the purpose and characteristics of domestic patents were Artificial Floating Island. In particular, domestic leisure space with a growing population and the need for securing emerging role as a reservoir of water only in the past, who do appeal as a tourist destination or as an ecological space utilized, and accordingly will transform and the need to secure a hydrophilic, degrade water quality problems using this aquatic environment (water acquisition and hydrophilic), the requirements are a big obstacle is the reality factor. This patented product differentiation strategy through the analysis of the development of technology progressiveness (Field Application) in terms of water quality improvement and maintenance side, and the hydrophilic side scenery, ecological restoration aspects, and applicability to the field and taking into account existing technology economic aspects of distinction were presented and advertised a lot in terms of cost compared to other techniques without the use of highly efficient methodology for building a water purification and also appears identity appeal, wetlands, rivers, etc. can be applied broadly technician widespread deployment and installation time to less simple and more are expected to spread.

기능화에 의한 단일벽 탄소나노튜브 정제 및 페이퍼 제조와 전계방출 특성 연구 (Preparation of Bucky Paper using Single-walled Carbon Nanotubes Purified through Surface Functionalization and Investigation of Their Field Emission Characteristics)

  • 곽정춘;이승환;이한성;이내성
    • 한국전기전자재료학회논문지
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    • 제21권5호
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    • pp.402-410
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    • 2008
  • Single-walled carbon nanotubes (SWCNTs) were currently produced together with some contaminants such as a metallic catalyst, amorphous carbon, and graphitic nanoparticles, which should be sometimes purified for their applications. This study aimed to develop efficient, scalable purification processes but less harmful to SWCNTs. We designed three-step purification processes: acidic treatment, surface functionalization and soxhlet extraction, and heat treatment. During the soxhlet extraction using tetrahydrofuran, specifically, carbon impurities could be easily expelled through a glass thimble filter without any significant loss of CNTs. Finally, SWCNTs were left as a bulky paper on the filter through membrane filtration. Vertically aligned SWCNTs on one side of bulky paper were well developed in a speparation from the filter paper, which were formed by being sucked through the filter pores during the pressurized filtration. The bucky paper showed a very high peak current density of field emission up to $200\;mA/cm^2$ and uniform field emission images on phosphor, which seems very promising to be applied to vacuum microelectronics such as microwave power amplifiers and x-ray sources.

도시가스 용 바이오 가스 정제 시스템 개발 (Development of Biogas Purification System for City Gas Supply)

  • 이현진;고상욱;이인동;정인희;고재욱
    • 한국가스학회지
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    • 제23권2호
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    • pp.61-67
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    • 2019
  • 우리나라는 천연가스 수입국으로 2018년 미국이 수출한 물량의 20%를 수입할 정도로 많은 양을 수입하고 있다. 이에 가스 수요를 만족시키며, 기후변화 대응에 효과적으로 대응할 수 있는 바이오가스는 대체제가 될 수 있을 것으로 생각된다. 그러나 바이오가스의 생산량의 20%만이 판매되고 있고, 이 역시 효율이 좋지 못해 활용하기 어려운 실정이다. 본 연구에서는 바이오가스를 도시가스로 공급 할 수 있는 최적의 정제 시스템을 개발을 목적으로 하였다. 시스템 선정을 위한 바이오가스에 대한 분석, 시스템 설계를 위한 사례 발굴, 시나리오 구성, 비용편익 툴을 개발하고 사례 적용하여 최적의 시스템을 개발하고자 하였다.

Microfluidic Components and Bio-reactors for Miniaturized Bio-chip Applications

  • Euisik Yoon;Yun, Kwang-Seok
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권2호
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    • pp.86-92
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    • 2004
  • In this paper miniaturized disposable micro/nanofluidic components applicable to bio chip, chemical analyzer and biomedical monitoring system, such as blood analysis, micro dosing system and cell experiment, etc are reported. This system includes various microfluidic components including a micropump, micromixer, DNA purification chip and single-cell assay chip. For low voltage and low power operation, a surface tension-driven micropump is presented, as well as a micromixer, which was implemented using MEMS technology, for efficient liquid mixing is also introduced. As bio-reactors, DNA purification and single-cell assay devices, for the extraction of pure DNA from liquid mixture or blood and for cellular engineering or high-throughput screening, respectively, are presented.

홍조류인 한국산 김종에서의 염색체 DNA 분리방법 (Rapid and Efficient Purification of Nucleic Acids from the Macroalga Porphyra(Rhodophyta))

  • 류태형;최학선;최경희;이춘환
    • 한국식품영양과학회지
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    • 제27권6호
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    • pp.1166-1172
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    • 1998
  • A method for the isolation and purification of DNA from a red algae, Porphyra was innovated. The innovation of the method consists mainly of three steps that include sodium acetate treatment, chloroform extraction, and 0.2 volume isopropanol precipitation step. The sodium acetate treatment was designed to remove polysaccharide contamination, and the isopropanol step to remove proteins and salts contaminents. Genomic DNA,s of several species(for example, P. tenera, P. yezoensis, P. seriata, and P. pseudolinearis) was successfully isolated by the innovated method. The amount of DNA purified from one g of sample material with the innovated method was 53 g in average. The resulting DNA was characterized to include high molecular weight and showed no nuclease activity. The DNA was pure enough to be digested directly by various restriction enzymes without any difficulties. Porphyra DNA was pure enough and adequate for amplification reaction through the polymerase chain reaction (small nuclear rDNA PCR amplification).

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Prepurification of paclitaxel by micelle and precipitation

  • 성주리;안희분;김진현
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XII)
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    • pp.501-504
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    • 2003
  • A novel prepurification method was developed aiming at increasing yield and purity, also reducing solvent usage for purification of paclitaxel. This method was a simple and efficient procedure, for the isolation and prepurification of paclitaxel from the biomass of Taxus chinensis, consisting of micelle formation, followed by two steps of precipitation. The use of a micelle and precipitation in the prepurification process allows for rapid separation of paclitaxel from interfering compounds and dramatically reduces solvent usage compared to alternative methodologies. This prepurification process serves to minimize the size and complexity of the HPLC operations for paclitaxel purification. This process is readily scalable to a pilot plant and eventually to a production environment where multikilogram quantities of material are expected to be produced. As much as possible, the process has been optimized to minimize solvent usage, complexity, and operating costs.

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Optimization of RNA Purification Method from Ecklonia cava Kjellman (Laminariales, Phaeophyceae)

  • Ahn, Jong-Sung;Woo, Seon-Ock;Kim, Jeong-Ha;Oh, Yoon-Sik;Oak, Jung-Hyun;Yum, Seung-Shic
    • ALGAE
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    • 제19권2호
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    • pp.123-127
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    • 2004
  • A more rapid and efficient method to extract RNA from Ecklonia cava Kjellman (Laminariales, Phaeophyceae) was introduced in this study. Each step of the procedure was evaluated and the optimal concentration of each chemical in the lysis solution was determined. Tissue pulverization with PVPP and β-mercaptoethanol in the lysis solution were not essential for RNA extraction of this species. The highest yield and purity of E. cava RNA were obtained by the lysis solution containing 1% CTAB, 1 M NaCl, 0.7% PVP, 10mM EDTA and 100mM Tris-Cl (pH 9.0). Approximately 8μg of RNA was obtained from 200 mg of ground tissue. The ratios of the absorbance at 260 nm and 280 nm were from 1.6 to 1.8 and those of at 230 nm and 260 nm were from 1.8 to 2.0. The extracted RNAs obtained in this study turned out to have a sufficient quality for cDNA synthesis.

A Simultaneous Design of TSK - Linguistic Fuzzy Models with Uncertain Fuzzy Output

  • Kwak, Keun-Chang;Kim, Dong-Hwa
    • 제어로봇시스템학회:학술대회논문집
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    • 제어로봇시스템학회 2005년도 ICCAS
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    • pp.427-432
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    • 2005
  • This paper is concerned with a simultaneous design of TSK (Takagi-Sugeno-Kang)-linguistic fuzzy models with uncertain model output and the computationally efficient representation. For this purpose, we use the fundamental idea of linguistic models introduced by Pedrycz and develop their comprehensive design framework. The design process consists of several main phases such as (a) the automatic generation of the linguistic contexts by probabilistic distribution using CDF (conditional density function) and PDF (probability density function) (b) performing context-based fuzzy clustering preserving homogeneity based on the concept of fuzzy granulation (c) augment of bias term to compensate bias error (d) combination of TSK and linguistic context in the consequent part. Finally, we contrast the performance of the enhanced models with other fuzzy models for automobile MPG predication data and coagulant dosing process in a water purification plant.

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