• Title/Summary/Keyword: gel permeation chromatography

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Studies on the Stability of $Methylenediphosphonate-^{99m}Tc$ (Methylenediphosphonate $(MDP)-^{99m}Tc$의 안정도(安定度)에 관(關)한 연구(硏究))

  • Kim, Jae-Rok;Awh, Ok-Doo;Park, Kwung-Bae;Koo, Hyeon-Sook
    • The Korean Journal of Nuclear Medicine
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    • v.16 no.2
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    • pp.81-84
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    • 1982
  • To evaluate the in-vivo and in-vitro stability of methylene diphosphonate $(MDP)-^{99m}Tc$, relative ligand exchange rates in phosphate buffer between $MDP-^{99m}Tc$ and human serum albumin (HSA), and between pyrophosphate$(PYP)-^{99m}Tc$ and HSA have been measured. Gel permeation chromatography has also been carried out to estimate relative rates of ligand exchange between polysaccharide and the $^{99m}Tc-bone$ agents. The in-vitro stability was further correlated with its specific radioactivity. The results indicated that the $MDP-^{99m}Tc$ is more stable than the $PYP-^{99m}Tc$ but less stable than $MIDA-^{99m}Tc$, $HIDA-^{99m}Tc$, and $DTPA-^{99m}Tc$ etc. in refering to other data. In stability point of view, $MDP-^{99m}Tc$ is considered to be a better bone agent than $PYP-^{99m}Tc$ since $MDP-^{99m}Tc$ can be accumulated at bone by a smooth transfer of $^{99m}Tc$ to hydroxyapatite, whereas in case of $PYP-^{99m}Tc$, most of the $^{99m}Tc$ is transferred to HSA before arriving to the hydroxyapatite.

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Production and Characterization of an Anti-Angiogenic Agent front Saccharomyces cerevisiae K-7

  • Jeong, Seung-Chan;Lee, Dae-Hyoung;Lee, Jong-Soo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1904-1911
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    • 2006
  • The cell-free extracts of 250 yeasts were screened for their in vitro anti-angiogenic activity, to develop a new cancer metastasis inhibitor. Saccharomyces cerevisiae K-7 was selected as the producer of the anti-angiogenic agent, because it had the highest anti-angiogenic activity. The anti-angiogenic agent was produced maximally from hydrolysates of Saccharomyces cerevisiae K-7, when the yeast was cultured in yeast extract-peptone-dextrose medium at 30$^{\circ}C$ for 24 h, and cell-free extracts were than digested with pepsin for 4 h at 37$^{\circ}C$. The anti-angiogenic agent was further purified by ultrafiltration, Sephadex G-25 gel permeation chromatography and reverse-phase HPLC, and the anti-angiogenic activity of the final purified preparation was 72.7% at 10 $\mu$M/egg. The purified anti-angiogenic agent was found to originate from the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) molecule of Saccharomyces cerevisiae K-7, and its peptide sequence was Val-Ser-Trp-Tyr-Asp-Asn-Glu-Tyr-Gly-Tyr-Ser-Thr-Arg-Val-Val-Asp. In the MTT assay, the shape of the HT-l 080 cell was clearly changed to a circular type at 0.2 mM purified anti-angiogenic agent. This result indicated that the growth of the HT-I080 cell was significantly inhibited at 0.2 mM of the purified anti-angiogenic agent. The MMP activity of the treated HT-l080 cells was not affected, evidenced by the gelatin zymography, indicating that the anti-angiogenic mechanism of the purified anti-angiogenic agent is not mediated through MMP activity.

Functional Expression of Amylosucrase, a Glucan-Synthesizing Enzyme, from Arthrobacter chlorophenolicus A6

  • Seo, Dong-Ho;Jung, Jong-Hyun;Choi, Hyun-Chang;Cho, Hyun-Kuk;Kim, Hee-Hang;Ha, Suk-Jin;Yoo, Sang-Ho;Cha, Jaeho;Park, Cheon-Seok
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1253-1257
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    • 2012
  • A gene (acas) designated as ${\alpha}$-amylase was cloned from Arthrobacter chlorophenolicus A6. The multiple amino acid sequence analysis and functional expression of acas revealed that this gene really encoded an amylosucrase (ASase) instead of ${\alpha}$-amylase. In fact, the recombinant enzyme exhibited typical ASase activity by showing both sucrose hydrolysis and glucosyltransferase activities. The purified enzyme has a molecular mass of 72 kDa and exhibits optimal hydrolysis activity at $45^{\circ}C$ and a pH of 8.0. The analysis of the oligomeric state of ACAS with gel permeation chromatography revealed that the ACAS existed as a monomer.

Isolation, Purification, and Enzymatic Characterization of Extracellular Chitosanase from Marine Bacterium Bacillus subtilis CH2

  • Oh, Chul-Hong;Zoysa, Mahanama De;Kang, Do-Hyung;Lee, Young-Deuk;Whang, Il-Son;Nikapitiya, Chamilani;Heo, Soo-Jin;Yoon, Kon-Tak;Affan, Abu;Lee, Je-Hee
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1021-1025
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    • 2011
  • A Bacillus subtilis strain was isolated from the intestine of Sebastiscus marmoratus (scorpion fish) that was identified as Bacillus subtilis CH2 by morphological, biochemical, and genetic analyses. The chitosanase of Bacillus subtilis CH2 was best induced by fructose and not induced with chitosan, unlike other chitosanases. The strain was incubated in LB broth, and the chitosanase secreted into the medium was concentrated with ammonium sulfate precipitation and purified by gel permeation chromatography. The molecular mass of the purified chitosanase was detected as 29 kDa. The optimum pH and temperature of the purified chitosanase were 5.5 and $60^{\circ}C$, respectively. The purified chitosanase was continuously thermostable at $40^{\circ}C$. The specific acitivity of the purified chitosanase was 161 units/mg. The N-terminal amino acid sequence was analyzed for future study.

Studies on the Preparation and the Properties of Chitosan Oligosaccharide and its Derivatives (키토산 올리고사카라이드 및 그 유도체의 제조와 특성에 관한 연구)

  • Ha, Byeong-Jo;Kim, Jun-O;Lee, Ok-Seop
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.23 no.2
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    • pp.48-62
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    • 1997
  • Chitosan oligosaccharides having aldehyde group at reducing end were prepared by oxidative-deamination reaction of chitosan by using sodium nitrite, and the resulting aldehyde group was reduced to 2, 5-anhydro-D-mannitol group. The obtained chitosan oligosaccharides showed an average degree of polymerization 2~3 by gel permeation chromatography analysis. It was highly soluble in hydrophilic solvents and thermally stable. N, N-diacyl, O-acyl chitosan oligosaccharides were obtained from the reaction between chitosan oligosaccharides and acyl chloried under dimethylaminopyridine. From differential scanning calorimetric measurement, N, N-dilauroyl, O-lauroyl chitosan oligosaccharides showed mesophase region, which was confirmed by polarized microscope as a thermotropic liquid crystalline state. X-ray diffraction pattern revealed that N, N-dilauroyl, O-lauroyl chitosan oligosaccharedes were highly crystalline, whereas chitosan oligosaccharides were not.

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Synthesis and Properties of Hyperbranched Polyester with Second-Order Outical Nonlinearity (2차 비선형 광학 초분지형 폴리에스테르의 합성 및 특성)

  • 이종협;이광섭
    • Polymer(Korea)
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    • v.25 no.6
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    • pp.803-810
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    • 2001
  • A nonlinear optical hyperbranched polyester (PE-Azo/Hyper) was synthesized from 4-[N,N-bis(hydroxyethyl)amino-4'-formyl] azobenzene and cyanoacetic acid by a Knoevenagel polycondensation using 4-(dimethylamino) pyridine as a base. The resulting polymer was soluble in polar aprotic solvents such as N,N-dimethylformamide and 1-methyl-2-pyrrolidinone and could be processed into optical quality films by spin coating. The molecular weight was determined to be $M_w$=61,800 ($M_W/M_n{=1.86}$) by gel permeation chromatography using polystyrene as a standard. No melting point was detected by differential scanning Calorimeter, indicating that this polymer presents an amorphous state. It shows a glass transition temperature of $121^{\circ}C$. The second-order nonlinear optical coefficient $d_{33}$ of the poled polymer determined by the second harmonic generation at 1064 nm was 25.4 pm/V.

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Synthesis of Polystyrene-b-Poly(ethylene oxide)-b-Polylactide Copolymers via Sequential Anionic and Ring-Opening Polymerizations (순차적 음이온 및 개환중합반응을 통한 폴리스티렌-폴리에틸렌옥사이드-폴리락티드 블록공증합체의 합성)

  • Song, Jie;Cho, Byoung-Ki
    • Polymer(Korea)
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    • v.33 no.5
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    • pp.458-462
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    • 2009
  • We have synthesized ABC linear triblock copolymers, i.e., polystyrene-b-poly(ethylene oxide)-b-polylactide, via sequential anionic and ring-opening polymerizations. In the first anionic polymerization step, styrene was polymerized in cyclohexane using sec-butyllithium as the initiator. Poly (styryl) lithium was hydroxylated by the addition of ethylene oxide, and the subsequent protonation with methanolic HCl. In the second anionic polymerization step, potassium naphthalenide was used to deprotonate the hydroxyl group of the PS to generate the macroinitiator of PS-$O^-K^+$. Polymerization of ethylene oxide was performed in THF and terminated with methanolic HCl. In the ring-opening polymerization step, the PS-b-PEO-$AlEt_2$ macroinitiator was prepared from an $AlEt_3$/pyridine system in THF, and the polymerization of lactide was performed at $90^{\circ}C$. The resulting block copolymers showed well-defined molecular weights and narrow molecular weight distributions as revealed by $^1H$- NMR spectroscopy and gel permeation chromatography (GPC).

Chemoenzymatic Synthesis of H-shaped Amphiphilic Pentablock Copolymer and Its Self-assembly Behavior (H-형태 양친매성 펜타블록 공중합체의 화학효소적 합성과 자기회합거동 평가)

  • Chen, Peng;Li, Ya-Peng;Li, Cai-Jin;Meng, Xin-Lei;Zhang, Bao;Zhu, Ming;Liu, Yan-Jing;Wang, Jing-Yuan
    • Polymer(Korea)
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    • v.37 no.3
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    • pp.332-341
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    • 2013
  • H-shaped amphiphilic pentablock copolymers $(PSt)_2-b-PCL-b-PEO-b-PCL-b-(PSt)_2$ was synthesized via chemoenzymatic method by combining enzyme-catalyzed ring-opening polymerization (eROP) of ${\varepsilon}$-caprolactone (${\varepsilon}$-CL) and atom transfer radical polymerization (ATRP) of styrene. By this process, we obtained copolymers with controlled molecular weight and low polydispersity. The structure and composition of the obtained copolymers were characterized by nuclear magnetic resonance (NMR), gel permeation chromatography (GPC) and infrared spectroscopy analysis (IR). The crystallization behavior of the copolymers was analyzed by differential scanning calorimetry (DSC) and X-ray diffraction (XRD). The crystallization behavior of the H-shaped block copolymers demonstrated a PCL dominate crystallization. The self-assembly behavior of the copolymers was investigated in aqueous media. The hydrodynamic diameters of the copolymer micelles in aqueous solution were measured by dynamic light scattering (DLS). The morphology of the copolymer micelles was observed by atomic force microscopy (AFM) and transmission electron microscopy (TEM). The hydrodynamic diameters of spherical micelles declined gradually with the increase of the hydrophobic chain lengths of the copolymers. The critical micelle concentration (CMC) values were determined from fluorescence emission, and it was found that the CMCs decreased with an increase of PSt hydrophobic block lengths.

Synthesis and Characterization of Energetic Thermoplastic Elastomers based on Carboxylated GAP Copolymers

  • Lim, Minkyung;Jang, Yoorim;Kweon, Jeong-Ohk;Seol, Yang-Ho;Rhee, Hakjune;Noh, Si-Tae
    • Applied Chemistry for Engineering
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    • v.31 no.3
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    • pp.284-290
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    • 2020
  • Energetic thermoplastic elastomers (ETPEs) based on glycidyl azide polymer (GAP) and carboxylated GA copolymers [GAP-ETPE and poly(GA-carboxylate)-ETPEs] were synthesized using isophorone diisocyanate (IPDI), dibutyltin dilaurate (DBTDL), 1,4-butanediol (1,4-BD), and soft segment oligomers such as GAP and poly(GA-carboxylate). The synthesized GAP-ETPE and poly(GA-carboxylate)-ETPEs were characterized by Fourier transform infrared (FT-IR), gel permeation chromatography (GPC), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), universal testing machine (UTM), calorimetry and sensitivity towards friction and impact. DSC and TGA results showed that the introduction of carboxylate group in GAP helped to have better thermal properties. Glass transition temperatures of poly(GA-carboxylate)-ETPEs decreased from -31 ℃ to -33 ℃ compared to that of GAP-ETPE (-29 ℃). The first thermal decomposition temperature in poly(GA0.8-octanoate0.2)-ETPE (242 ℃) increased in comparison to that of GAP-ETPE (227 ℃). Furthermore, from calorimetry data, poly(GA-carboxylate)-ETPEs exhibited negative formation enthalpies (-6.94 and -7.21 kJ/g) and higher heats of combustion (46713 and 46587 kJ/mol) compared to that of GAP-ETPE (42,262 kJ/mol). Overall, poly(GA-carboxylate)-ETPEs could be good candidates for a polymeric binder in solid propellant due to better energetic, mechanical and thermal properties in comparison to those of GAP-ETPE. Such properties are beneficial to application and processing of ETPE.

Possible Presence of an Interleukin-6-Like Molecule in the Immunized Bombyx mori L. (Lepidoptera)

  • Kim, Iksoo;Lee, Young-Shin;Lee, Joon-Ha;Kim, Sang-Hyun;Kang, Pil-Don;Lee, In-Hee;Kim, Jin-Won;Lee, Heui-Sam;Kang, Seok-Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.7 no.2
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    • pp.165-173
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    • 2003
  • Cytokines represent an essential part of the innate immune response in mammals. Recently, several studies have reported the presence of cytokine-like activities and molecules in the invertebrates such as echinoderms, tunicates, mollusks and insects. In our serial study, we investigated presence of cytokines in the silkworm, Bombyx mori, infected with several immune inducers. Western blotting analysis using rabbit anti-human cytokines showed the presence of IL-6-like molecule in the hemolymph collected at 8 and 24 hrs after infection with peptidoglycan and oligodeoxynucleotide, and the molecular weight of the proteins was ∼45 kDa. We attempted to isolate the molecule by gel permeation HPLC, anion exchange chromatography, ultra centrifugation, and immuno-dot-blot assay, but until now the effort was not much successful yet. It, however, does not appear that the IL-6-like molecule in the silkworm larvae is a mere experimental artifact happened by Western blotting analysis. Instead, further experiment on this subject probably will provide us more fruitful result as detected in other invertebrates including insects.