• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.4
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• pp.263-267
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• 2003

Magnetic nanoparticles prepared from an alkaline solution of divalent and trivalent iron ions could covalently bind protein via the activation of Nethyl-N-(3-dimethylaminopropyl) carbodiimide (EDC). Trypsin and avidin were taken as the model proteins for the formation of protein-nanoparticle conjugates. The immobilized yield of protein increased with molar ratio of EDC/nanoparticie. Higher concentrations of added protein could yield higher immobilized protein densities on the particles. In contrast to EDC, the yields of protein immobilization via the a ctivation of cyanamide were relatively lower. Nanoparticles bound with avidin could attach a single-stranded DNA through the avidin-biotin interaction and hybridize with a DNA probe. The DNA hybridization was confirmed by fluorescence microscopy observations. Immobilized DNA on nanoparticles by this technique may have widespread applicability to the detection of specific nucleic acid sequence and targeting of DNA to particular cells.

• Advances in nano research
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• v.6 no.1
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• pp.81-92
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• 2018

Biosynthesis of nanoparticles has acquired particular attention due to its economic feasibility, low toxicity and simplicity of the process. Extracellular synthesis of nanoparticles by bacteria and fungi has been stated to be brought about by enzymes and other reducing agents that may be secreted in the culture medium. The present study was carried out to determine the underlying mechanisms of extracellular silver nanoparticle synthesis by Pseudomonas hibiscicola isolated from the effluent of an electroplating industry in Mumbai. Synthesized nanoparticles were characterized by spectroscopy and electron microscopic techniques. Protein profiling studies were done using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (1D-SDS PAGE) and subjected to identification by Mass Spectrometry. Characterization studies revealed synthesis of 50 nm nanoparticles of well-defined morphology. Total protein content and SDS PAGE analysis revealed a reduction of total protein content in test (nanoparticles solution) samples when compared to controls (broth supernatant). 45.45% of the proteins involved in the process of nanoparticle synthesis were identified to be oxidoreductases and are thought to be involved in either reduction of metal ions or capping of synthesized nanoparticles.

• Animal cells and systems
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• v.15 no.2
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• pp.107-114
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• 2011

A proteomic analysis of the proteins in mouse brain that were differentially expressed in response to $TiO_2$ nanoparticles was conducted to better understand the molecular mechanism of $TiO_2$ nanoparticle-induced brain toxicity at the protein level. A total of 990 proteins from mouse brain were resolved by two-dimensional gel electrophoresis. A comparative proteomic analysis revealed that the expression levels of 11 proteins were changed by more than 2-fold in response to $TiO_2$ nanoparticles: eight proteins were upregulated and three were downregulated by $TiO_2$ nanoparticles. In addition, the activities of several antioxidative enzymes and acetylcholine esterase were reduced in $TiO_2$ nanoparticle-exposed mouse brain. The protein profile alterations seem to be due to an indirect effect of $TiO_2$ nanoparticles, because $TiO_2$ nanoparticles were not detected in the brain in this investigation.

• Bulletin of the Korean Chemical Society
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• v.31 no.2
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• pp.479-482
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• 2010

• Proceedings of the Korean Vacuum Society Conference
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• 2015.08a
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• pp.230.1-230.1
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• 2015

One of the major problems of biological ToF-SIMS imaging is the lack of protein and peptide imaging. Most of biological story telling is mianly based on proteins. The biological implication of lipid ToF-SIMS imaging would be much higher if protein imaging is provided together. Utilizing high secondary ion yields of metals, proteins can be ToF-SIMS imaged with nanoparticle tagged proteins. Nanoparticles such as Fe3O4, SiO2, PbS were used for imaing NeuN, MCH, Orexin A, ${\alpha}$ synucline, TH(Tryosine Hydroxylase) in mouse tissues with a spatial resolution of ${\sim}2{\mu}m$ using a TOF-SIMS. Lipids and neurotransmitters images obtained simultaneously with protein images were overlayed for more deeper understanding of neurobiology, which is not allowed by any other bioimaging technqiues. The protein images from TOF-SIMS were compared with confocal fluorescence microscopy and NanoSIMS images. A new sample preparation method for imaging single cell membranes in a tissue using the vibrotome technique to prepare a tissue slice without any fixation and freeze drying will be also presented briefly for Hippocampus and Hypothalamus tissues.

• BMB Reports
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• v.55 no.4
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• pp.175-180
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• 2022

Peptides are gaining substantial attention as therapeutics for human diseases. However, they have limitations such as low bioavailability and poor pharmacokinetics. Periostin, a matricellular protein, can stimulate the repair of ischemic tissues by promoting angiogenesis. We have previously reported that a novel angiogenic peptide (amino acids 142-151) is responsible for the pro-angiogenic activity of periostin. To improve the in vivo delivery efficiency of periostin peptide (PP), we used proteins self-assembled into a hollow cage-like structure as a drug delivery nanoplatform in the present study. The periostin peptide was genetically inserted into lumazine synthase (isolated from Aquifex aeolicus) consisting of 60 identical subunits with an icosahedral capsid architecture. The periostin peptide-bearing lumazine synthase protein cage nanoparticle with 60 periostin peptides multivalently displayed was expressed in Escherichia coli and purified to homogeneity. Next, we examined angiogenic activities of this periostin peptide-bearing lumazine synthase protein cage nanoparticle. AaLS-periostin peptide (AaLS-PP), but not AaLS, promoted migration, proliferation, and tube formation of human endothelial colony-forming cells in vitro. Intramuscular injection of PP and AaLS-PP increased blood perfusion and attenuated severe limb loss in the ischemic hindlimb. However, AaLS did not increase blood perfusion or alleviate tissue necrosis. Moreover, in vivo administration of AaLS-PP, but not AaLS, stimulated angiogenesis in the ischemic hindlimb. These results suggest that AaLS is a highly useful nanoplatform for delivering pro-angiogenic peptides such as PP.

• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.501-519
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• 2003

Silk Sericin(SS) is a natural protein extracted from cocoon of bombix mori and shows moisturizing effect to the skin due to a number of hydroxyl groups in the structure. But its application to cosmetics is limited due to its poor solubility in water. In order to solve this drawback and expand its application to cosmetics, polyethyleneglycol(PEG) was conjugated with sericin by reacting activated polyethyleneglycol(ActPEG). Reaction site of sericin is tyrosine residue, which was determined by using $^1$H-NMR. Random coil structure of sericin was transformed to beta-sheet structure by conjugating polyethyleneglycol. It was confirmed that melting point of sericin-PEG conjugate was lowered compared to that of each sericin and PEG due to the interaction between sericin and PEG in crystalline structure. Self-assembled sericin-PEG nanoparticle was obtained by dialyzing with alcohol solution of sericin-PEG conjugate against water. The particle is spherical and has 200-400nm of size. The moisturizing ability of sericin-PEG nanoparticle was much higher than that of sericin itself. Incorporation of vitamin A into sericin-PEG nanoparticle was carried out by diafiltration method. The content of incorporated Vitamin A in sericin-PEG nanoparticle was 8.9 wt％. Releasing behaviour of vitamin A incorporated into nanoparticle was tested in phosphate buffer, pH 7.4 at 37$^{\circ}C$. and half-life of Vitamin A release was 43hrs. Sericin-PEG nanoparticle exhibited higher moisturing effect than sericin itself and distilled water, respectively. No toxicity and irritation were observed in animal tests. It can be expected that the self-assembled sericin-PEG nanoparticle can be developed for cosmetics.

• Proceedings of the Materials Research Society of Korea Conference
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• 2003.03a
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• pp.51-51
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• 2003

• 한국당과학회:학술대회논문집
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• 2006.11a
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• pp.15-15
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• 2006

• Polymer(Korea)
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• v.32 no.3
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• pp.246-250
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• 2008

Colloidal stability of the biodegradable nanoparticle was characterized by measuring the variation of particle size with time using photon correlation spectroscopy. Three kinds of polymers, namely, poly(D,L-lactide-co-glycolide)(PLGA), PLGA/poly(L-lactide) blends, and PLGA/poly(L-lactide)-g-poly(ethylene glycol) blends were used as matrix material for nanoparticle preparation. Nanoparticles were prepared with or without using poly(vinyl alcohol)(PVA) as suspension stabilizer to evaluate the condition of preparation. Nanoparticles from the blend of amphiphilic graft copolymer with short poly(ethylene glycol) chain and PLGA maintained suspension for 1 day when protein stock solution was introduced. This is somewhat improvement in colloidal stability against protein adsorption compared with that of nanoparticles without PEG moiety. Suspension stabilizer, PVA, had a significant effect on the colloidal stability against freezing and protein adsorption which led to coagulation of nanoparticles. It is important to consider effect of suspension stabilizer as well as materials used to prepare nanoparticle on the colloidal stability.