• Biomedical Science Letters
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• v.12 no.3
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• pp.177-183
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• 2006

To demonstrate the effect of formulation conditions and evaluations of structural integrity from ovalbumin containing poly lactide glycolide copolymer (PLGA) microspheres for Vaccine delivery, OVA microspheres were prepared by a W/O/W multiple emulsion solvent extraction technique. Dichloromethan (DCM) and Ethyl acetate (EA) were applied as an organic phase and poly vinyl alcohol (PVA) as a secondary emulsion stabilizer. Microspheres were characterized for particle size, morphology (optical microscopy and Scanning Electron Microscope (SEM)). Protein denaturation was evaluated by size exclusion chromatography (SEC), SDS-PAGE and isoelectric focusing (IEF). Residual organic solvent was estimated by gas chromatography (GC) and differential scanning calorimetry (DSC). Optical photomicrograph and SEM revealed that micro spheres were typically spherical but various morphologies were observed. Mean particle size $(d_{vs})$ of microspheres were in the range of $3{\sim}50{\mu}m$. Also, The protein stability was not affected by the fonnulation process and residual organic solvent was beyond the detection below 0.1ppm. These results demonstrated that micro spheres might be a good candidate for the parenteral vaccine delivery system.

• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1250-1257
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• 2005

Two groups of exopolysaccharides (designated as Fr-I EPS and Fr-II EPS) were isolated from the culture filtrate of new fungal strain Trichoderma erinaceum DG-312 by Sepharose CL-6B chromatography. The structures of the exopolysaccharides were investigated using gas chromatography (GC), Fourier transform-infrared (FT-IR) spectroscopy, GCMS analysis, and NMR. GC analysis indicated that Fr-I EPS was composed of mainly mannose ($78.9\%$) and galactose ($21.1\%$), whereas Fr-II EPS contained mannose ($68.4\%$), galactose ($26.2\%$), and glucose ($5.4\%$). In the anomeric region ($950-700cm_{-1}$) of the FT-IR spectrum, both EPSs exhibited obvious characteristic absorption of $810\;cm_{-1}$, indicating the existence of mannose. The spectra of $\alpha-and\;\beta$-configurations were assigned at 880 and $914\;cm_{-1}$, respectively. The results of GC-MS analyses confirmed that both EPSs were complex heteropolysaccharides with a ($1{\rightarrow}3$)-linked mannan backbone. The C-1 region that appeared in the $^{13}C-NMR$ spectra of these EPSs indicated a typical anomeric carbon signal. The Fr-I EPS showed two anomeric carbon signals at 102.6 and 99.6 ppm, whereas the Fr-II EPS displayed four anomeric carbon signals at 102.5, 99.6, 98.5, and 94.3 ppm. The molecular characteristics of the EPSs were further investigated using a size exclusion chromatography/multi-angle laser light scattering (SEC/MALLS) system. The SEC/MALLS system revealed that the average molar masses of the EPSs were $6.592{\times}10^{4}$ (Fr-I EPS) and $1.920{\times}10^{4}$ (Fr-II EPS) g/mol, and the molecular conformation of both EPSs in aqueous solution was random coils.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.11
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• pp.1054-1062
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• 2010

Oxidative coupling reactions of humic substances (HS) can be catalyzed by a variety of natural extracellular enzymes and metal oxides. In this study, property changes of HS induced by a natural enzyme, horseradish peroxidase (HRP), and the effect of it to microfiltration (MF) were investigated. PAHA was transformed by oxidative coupling reaction with HRP and hydrogen peroxide ($H_2O_2$), verifying the catalytic effects of the HRP. Size exclusion chromatography (SEC) revealed that weight-average molecular weight (MWw) of PAHA was proportionally increased with the dosages of HRP and $H_2O_2$, indicating the transform action of HS into larger and complex molecules. An increase in the conformational stability of HS was achieved through the promotion of intermolecular covalent bondings between heterogeneous humic molecules. Spectroscopic analysis (fluorescence and infrared spectroscopy) proved that functional groups were transformed by the reaction. Additionally, HS and transformed products were undergone microfiltration (MF) to examine the treatment potential of them in a water treatment facility. Original HS could not be removed by MF but larger molecules of transformed products could be removed. Meanwhile, transformed products caused more fouling on the filtration than original HS. This results proved that natural organic matter (NOM) can be removed by MF after its increase in molecular size by oxidative coupling reaction.

• Bulletin of the Korean Chemical Society
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• v.34 no.9
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• pp.2663-2670
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• 2013

The purpose of stability testing is to provide evidence about how the quality of a drug varies with time under the influence of a variety of environmental factors. In this study, erythropoietin (EPO) was analyzed under different pH (pH 3 and pH 9) and temperature ($25^{\circ}C$ and $40^{\circ}C$) conditions according to current Good Manufacturing Practice (cGMP) and International Conference on Harmonisation (ICH) guidelines. The molecular weight difference between intact EPO and deglycosylated EPO was determined by SDS-PAGE, and aggregated forms of EPO under thermal stress and high-pH conditions were investigated by size exclusion chromatography. High pH and high temperature induced increases in dimer and high molecular weight aggregate forms of EPO. UPLC-ESI-TOF-MS was applied to analyze the changed modification sites on EPO. Further, normal-phase high-performance liquid chromatography was performed to identify proposed glycan structures and high pH anion exchange chromatography was carried out to investigate any change in carbohydrate composition. The results demonstrated that there were no changes in modification sites or the glycan structure under severe conditions; however, the number of dimers and aggregates increased at $40^{\circ}C$ and pH 9, respectively.

• Journal of the Korean Chemical Society
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• v.42 no.2
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• pp.190-196
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• 1998

A new sampling and size exclusion chromatography (SEC) method for the analysis of underivatized cellulose are established. In this method, cellulose materials are first dissolved in N-methylmorpholine N-oxide (NMMO) and diluted by adding dimethyl sulfoxide (DMSO) to make the sample solutions of about 0.1% in 50/50 NMMO/DMSO (w/w). Sample solutions are analyzed using a glucose-treated divinylbenzene (DVB) SEC column and DMSO containing 0.05M LiBr and 2.5 blank as the eluant. The flow rate was constant at 1 mL/min and the whole SEC system including the column was heated at $80^{\circ}C$ to reduce the viscosity of DMSO. Addition of 0.05 M LiBr eliminated SEC baseline drifting, and addition of 2.5 blank seems to reduce the interaction between the sample and the column packing. SEC molecular weights were determined using a calibration curve constructed from a series of narrow pullulan standards, and they were used to measure the degree of degradation during two different pulp-to-sponge processings.

• Journal of Microbiology and Biotechnology
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• v.29 no.2
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• pp.274-282
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• 2019

Mercury-resistant ($Hg^R$) bacteria were isolated from heavy metal polluted wastewater and soil collected near to tanneries of district Kasur, Pakistan. Bacterial isolates AZ-1, AZ-2 and AZ-3 showed resistance up to $40{\mu}g/ml$ against mercuric chloride ($HgCl_2$). 16S rDNA ribotyping and phylogenetic analysis were performed for the characterization of selected isolates as Bacillus sp. AZ-1 (KT270477), Bacillus cereus AZ-2 (KT270478) and Bacillus cereus AZ-3 (KT270479). Phylogenetic relationship on the basis of merA nucleotide sequence confirmed 51-100% homology with the corresponding region of the merA gene of already reported mercury-resistant Gram-positive bacteria. The merE gene involved in the transportation of elemental mercury ($Hg^0$) via cell membrane was cloned for the first time into pHLV vector and transformed in overexpressed C43(DE3) E. coli cells. The recombinant plasmid (pHLMerE) was expressed and the native MerE protein was obtained after thrombin cleavage by size exclusion chromatography (SEC). The purification of fusion/recombinant and native protein MerE by Ni-NTA column, dialysis and fast protein liquid chromatography (FPLC/SEC) involved unfolding/refolding techniques. A small-scale reservoir of wastewater containing $30{\mu}g/ml$ of $HgCl_2$ was designed to check the detoxification ability of selected strains. It resulted in 83% detoxification of mercury by B. cereus AZ-2 and B. cereus AZ-3, and 76% detoxification by Bacillus sp. AZ-1 respectively (p < 0.05).

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.158-162
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• 2005

Enterobacteria, named ${\lambda}-bacteria$ isolated from fusiform fish, have strong anti-angiogenesis effect. ${\lambda}-28$ species bore higher anti-angiogenesis effect. Cultured liquid was performed salting out, dialysed and freezed dried. This sample was executed size exclusion chromatography with fraction collector. Anti-angiogenesis, cytotoxicity, and SDS-PAGE were carried out with fraction number. ${\lambda}-28$ species was lower toxicity against HUVECs and effective band was conformed with SDS-PAGE.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.43 no.5
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• pp.60-67
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• 2011

Hemicelluloses pre-extracted from Korean mixed wood chip were investigated as a wet-end additive. Hemicelluloses dissolved in hot water and alkali solution were isolated by ethyl alcohol precipitation from pre-extractives. They showed molecular weight of 9,000 ~ 27,000 g/mol as revealed by size exclusion chromatography. The reduction of molecular weight through hot water extraction was caused by autohydrolysis. Chemical composition of the hemicelluloses were analyzed with high-performance liquid chromatography and UV-Vis spectroscopy. As the surface charge of isolated hemicelluloses were negative, the adsorption of hemicelluloses onto softwood unbleached kraft pulp fiber was promoted by poly-DADMAC. The physical properties of handsheets increased as the molecular weight of hemicellulose increased. On the other hands, the optical property decreased with hemicellulose adsorption.

• Journal of Korean Society of Environmental Engineers
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• v.36 no.7
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• pp.492-497
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• 2014

This study investigated removal characteristics of organic matters in pretreatment and reverse osmosis (RO) membrane processes for seawater desalination. Also, the influence of the changes in characteristics of organic matters on the membrane fouling was assessed. The pretreatment processes included dual media filtration (DMF), pressurized membrane filtration (MF), and submerged membrane filtration (SMF). Turbidity, UV absorption at 254 nm, dissolved organic carbon, size exclusion chromatography (SEC), fluorescence excitation emission matrix (FEEM), and transparent exopolymer particles (TEP) in raw and processed waters were analyzed. Ions and minerals were not removed by any pretreatment process tested, but were removed over 99% through the RO membrane process. Hydrophobic organics, which can play major role in organic membrane fouling, were relatively readily removed compared with hydrophilic ones. Membrane based pretreatment such as MF and SMF exhibited better removals of organics than conventional DMF. As the levels of organics in pretreated water decreased, the silt density index (SDI) decreased. MF treated water exhibited the lowest SDI value; this is possibly due to the lowest TEP ($0.1-0.4{\mu}m$) concentrations.

• Journal of Korean Society on Water Environment
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• v.38 no.1
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• pp.31-42
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• 2022

Dissolved organic matter (DOM), which changes according to various factors, is ubiquitously present from natural environments to engineered treatment systems. Only limited information is available regarding the environmental functions of DOM after bulk analyses are only applied for characterization. In this paper, latest DOM analytical techniques are briefly introduced, which include fluorescence excitation-emission matrix with parallel factor analysis (EEM-PARAFAC), size-exclusion chromatography with an organic carbon detector (SEC-OCD), carbon/nitrogen stable-isotope ratio, and Fourier transform-ion cyclotron resonance-mass spectroscopy (FT-ICR-MS). Recent examples of using advanced analyses to interpret the phenomena associated with DOM occurring in natural and engineered systems are presented here. Through EEM-PARAFAC, different components like protein-like, fulvic-like, and humic-like can be identified and tracked individually through the investigated systems. SEC-OCD allows researchers to quantify different size fractions. FT-ICR-MS provides thousands of molecular formulas present in bulk DOM samples. Lastly, carbon/nitrogen stable-isotope ratio offers reasonable tools for tracking the sources in environments. We also discuss the advantages and weakness of the above-mentioned characterizing tools. Specifically, they focus on single environmental factors (different sourced-DOM and interaction of sediment-pore water) or simple changes after individual treatment processes. Through collaboration with the advanced techniques later, they help the researchers to better understand environmental behaviors in aquatic systems and serve as essential tools for addressing various pending problems associated with DOM.