• Title/Summary/Keyword: membrane chromatography

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Preparation of Polyvinylpyrrolidone/AgBF4/Al(NO3)3 Electrolyte Membranes for Facilitated Gas Transport (기체 촉진수송을 위한 polyvinylpyrrolidone/AgBF4/Al(NO3)3 전해질 분리막 제조)

  • Yoon, Ki Wan;Kang, Sang Wook
    • Membrane Journal
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    • v.26 no.1
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    • pp.38-42
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    • 2016
  • Polyvinylpyrrolidone (PVP), which is glassy polymer to have amide functional group, was induced to fabricate the facilitated olefin transport membranes for olefin/paraffin separation. Separation performance for the mixed gas consisting of propylene and propane (50 : 50 vol%) was measured by gas chromatography and bubble flow meter. The properties of membranes were confirmed by scanning electron microscope and FT-IR. The results of long-term separation tests showed the selectivity of 15 and permeance of 1.3 GPU. The membranes was compared with poly(2-ethyl-2-oxazoline) $(POZ)/AgBF_4/Al(NO_3)_3$ membranes and the characteristics were confirmed as polymer matrix for facilitated transport membranes.

Application of Membrane Modules to Affinity Chromatography Column (막모듈의 친화성 크로마토그래피 컬럼으로의 활용)

  • 이광진;김민정;염경호
    • Proceedings of the Membrane Society of Korea Conference
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    • 1998.04a
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    • pp.120-123
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    • 1998
  • 1. 서론 : 생물기술이 급속히 발전함에 따라 다양한 생물제품들(예, 단백질, 효소, 항생제 등)이 생산되고 있으나, 그 생산량은 극단적으로는 기질용액 1g당 10$^{-8}$g 정도로까지 낮아, 이를 고순도로 정제하기 위해서는 다단계의 공정이 필요하여 분리정제의 비용이 크다. 따라서 생물제품의 산업적 생산이 경제성을 갖기 위해서는 생물제품을 보다 경제적으로 분리정제할 수 있는 효율적인 공정개발이 필수적이다. (생략)

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Identification of Phospholipase C Activated by $GTP{\gamma}S$ in Plasma Membrane of Oat Cell

  • Kim, Hyae-Kyeong;Park, Moon-Hwan;Chae, Quae
    • BMB Reports
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    • v.28 no.5
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    • pp.387-391
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    • 1995
  • In order to investigate whether phospholipase C (PLC) activity in oat celIs is regulated by Gprotein, we have characterized PLC in plasma membranes of oat tissues. To identify the purified plasma membrane, $K^+$-stimulated, $Mg^{2+}$-dependent ATPase activity was measured. The activity of ATPase was shown to be proportional to the concentration of membrane protein. To examine the PLC activity regulated by G-protein, we used the inside-out and outside-out plasma membrane mixture isolated from the oat cells. The plasma membrane mixture showed higher PLC activity than the one of the outside-out plasma membrane. This suggests that PLC activity is located at the cytoplasmic surface of plasma membrane. PLC activity in plasma membrane mixture was dependent on $Ca^{2+}$ with maximum activity at 100 ${\mu}m$ $Ca^{2+}$ and it was inhibited by 1 mM EGTA. Using Sep-pak $Accell^{TM}$ Plus QMA chromatography, we found that inositol 1,4,5-trisphosphate ($IP_3$) was produced in the presence of 10 ${\mu}m$ $Ca^{2+}$. The PLC activity in the membrane was enhanced by an activator of G-protein ($GTP{\gamma}S$) and not by an inhibitor ($GDP{\beta}S$). This indicates that a G-protein is involved in the activation of PLC in the plasma membrane of oat cells.

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Isolation of GTP Binding Protein from Bovine Brain (소의 뇌로부터 GTP 결합단백질의 분리)

  • Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
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    • v.10 no.2
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    • pp.360-368
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    • 1993
  • GTP binding protein (G-protein) associated with membrane and involved in signal transduction was isolated from bovine brain, and molecular weight of G protein was observed. As the results, cell membranes were homogenized from bovine brain tissues and proteins of membrane were gained using 1% cholate, and progressed the chromatography. The purification process was performed by step, DEAE-Sephacel, Ulttrogel AcA 34 and heptylamine-Sepharose column chromatography. The chromatographic fractions were confirmed by GTP binding assay and SDS-polyacrylamide gel electrophoresis. Molecular weight of $Go{\alpha}$ was revealed 39,000 dalton and $G{\beta}$ 36,000 dalton. One more step of heptylamine-Sepharose was enforced to purify the GTP binding protein. Finally I gained the GTP binding protein isolated subtype of $Go{\alpha}$ and $G{\beta}$.

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Expression and Purification of a Cathelicidin-Derived Antimicrobial Peptide, CRAMP

  • Park Eu-Jin;Chae Young-Kee;Lee Jee-Young;Lee Byoung-Jae;Kim Yang-Mee
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1429-1433
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    • 2006
  • Application of recombinant protein production and particularly their isotopic enrichment has stimulated development of a range of novel multidimensional heteronuclear NMR techniques. Peptides in most cases are amenable to assignment and structure determination without the need for isotopic labeling. However, there are many cases where the availability of $^{15}N$ and/or $^{13}C$ labeled peptides is useful to study the structure of peptides with more than 30 residues and the interaction between peptides and membrane. CRAMP (Cathelicidin-Related AntiMicrobial Peptide) was identified from a cDNA clone derived from mouse femoral marrow cells as a member of cathelicidin-derived antimicrobial peptides. CRAMP was successfully expressed as a GST-fused form in E. coli and purified using affinity chromatography and reverse-phase chromatography. The yield of the CRAMP was 1.5 mg/l 1. According to CD spectra, CRAMP adopted ${\alpha}$-helical conformation in membrane-mimetic environments. Isotope labeling of CRAMP is expected to make it possible to study the structure and dynamic properties of CRAMP in various membrane systems.

Composite Membrane Containing a Proton Conductive Oxide for Direct Methanol Fuel Cell

  • Peck, Dong-Hyun;Cho, Sung-Yong;Kim, Sang-Kyung;Jung, Doo-Hwan;Kim, Jeong-Soo
    • Journal of the Korean Electrochemical Society
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    • v.11 no.1
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    • pp.11-15
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    • 2008
  • The composite membrane for direct methanol fuel cell (DMFC) was developed using $H_3O^+-{\beta}"-Al_2O_3$ powder and perfluorosulfonylfluroride copolymer (Nafion) resin. The perfluorosulfonylfluroride copolymer (Nafion) resin was mixed with $H_3O^+-{\beta}"-Al_2O_3$ powder and it was made to sheet form by hot pressing. The electrodes were prepared with 60 wt% PtRu/C and 60wt% Pt/C catalysts for anode and cathode, respectively. The morphology and the chemical composition of the composite membrane have been investigated by using SEM and EDXA, respectively. The composite membrane and $H_3O^+-{\beta}"-Al_2O_3$ were analyzed by using FT-IR and XRD. The methanol permeability of the composite membranes was also measured by gas chromatography (GC). The performance of the MEA containing the composite membrane (2wt% $H_3O^+-{\beta}"-Al_2O_3$) was higher than that of normal pure Nafion membrane at high operating temperature (e.g. $110^{\circ}C$), due to the homogenous distribution of $H_3O^+-{\beta}"-Al_2O_3$, which decreased the methanol permeability through the membrane and enhanced the water contents in the composite membrane.

Development of Immuno-Analytical System for Microbial Cells by using Dot-Blotter (Dot-Blotter 진공 포획방식에 의한 미생물세포 면역분석시스템의 개발)

  • 목락선;하연철;윤희주;백세환
    • KSBB Journal
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    • v.14 no.1
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    • pp.82-90
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    • 1999
  • In order to eventually fabricate an analytical system for infectious microorganisms, we synthesized major immunochemical components, utilized them for the construction of model system, and investigated an assay concept for bacterial whole cells. For the preparation of system components, a polyclonal antibody, against Salmonella thompson as model analyte, purified by immuno-affinity chromatography was used to chemically link to streptavidin or an enzyme, horseradish peroxidase(HRP). The antibody and streptavidin was modified with sulfosuccinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxylate and N-succinimidyl-3-[2-pyridyldithio]propionate(subsequently activated by dithiotheritol), respectively. The modified components were reacted to synthesize antibody-streptavidin conjugates which were then purified on a two-layer chromatography column of diaminobiotin gel and Sephadex G-100. For antibody-HRP conjugates, HRP molecules were activated by $NalO_4$ oxidation and then coupled to immunoglobulin. After stabilizing with ($NaCNBH_3$, the conjugates were purified by size exclusion chromatography on Biogel A5M column. To devise a model system, such produced components were combined with a dot-blotter in which a nitrocellulose membrane($12{\mu}m$ pre size) with immobilized biotin was already located. The analyte (S. thompson cells) was reacted with the both antibody conjugates in a liquid phase, and the complexes formed were captured on the membrane surfaces by applying vacuum in the bottom compartment of the blotter to invoke biotin-streptavidin reaction. Under optimal conditions, the system enabled to identify the analytical concept for bacterial whole cells, and the lower limit of detection was approximately $1{\mu}g/m{\ell}$($10^5-10^6$ cells/m$m{\ell}$). The controlling factors were the concentrations of each antibody conjugate that caused agglutination in the presence of analyte as they increased.

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Purification and Characterization of Recombinant Monellin Produced from Yeast Culture Medium (효모 배양액으로부터 재조합 모넬린의 정제와 특성 연구)

  • 김인호
    • KSBB Journal
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    • v.13 no.5
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    • pp.535-539
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    • 1998
  • The monellin, a sweet-taste protein, was expressed and secreted in Saccharomyces cerevisiae. The secreted menellin was concentrated using an ultrafiltration membrane with a nominal molecular weight cut off of 3,000 or by ammonium sulfate precipitation. The monellin was purified by G-25 gel filtration chromatography, followed by CM-Sepharose ion exchange chromatography. The purified monellin was characterized by SDS-PAGE (SDS-Polyacrylamide Gel Electrophoresis) and PHLC. The molecular weight of monellin was found to be 10,700 dalton, and its purity was over 95%.

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Analysis of the Limonoid Contents of Dangyuja (Citrus spp.) by Liquid Chromatography-mass Spectrometry (LC-MS) (Liquid chromatography-mass spectrometry(LC-MS)에 의한 당유자 과실의 리모노이드 함량 분석)

  • Boo, Kyung-Hwan;Kim, Ha-Na;Riu, Key-Zung;Kim, Youn-Woo;Cho, Moon-Jae;Kim, So-Mi
    • Applied Biological Chemistry
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    • v.50 no.3
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    • pp.238-243
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    • 2007
  • The contents of limonin, nomilin and limonin glucoside in different tissues of dangyuja (Citrus spp.) were measured during fruit growth and maturation by liquid chromatography-mass spectrometry(LC-MS). Both nomilin and limonin contents increased from June, peaked in October and in December, respectively, and then decreased afterwards. In contrast, the content of limonin glucoside increased from October and remained at high levels throughout the maturation. The contents of limonin, nomilin and limonin glucoside in peel decreased gradually throughout the growth and the trends of changes were similar to each other. However, all three limonoid contents in juice sac increased dramatically at the end of fruit maturation. The highest concentration of limonin and nomilin was observed in seed, followed by segment membrane, whereas the highest limonin glucoside concentration was observed in segment membrane, followed by juice sac.

Chromatography separation of proteins by macroporous chitosan and chitin affinity membranes

  • Yuk, Yeong-Jae;Youm, Kyung-Ho
    • Proceedings of the Membrane Society of Korea Conference
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    • 2004.05a
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    • pp.59-62
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    • 2004
  • Affinity membranes have emerged principally to overcome the problems of limited specificity experienced with membranes that operate purely on a sieving mechanism and as an alternative to the traditional affinity resins. It is a logical expectation that affinity membranes might combine the outstanding selectivity of affinity resins with the high productivity associated with filtration membranes.(omitted)

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