• The Korean Journal of Zoology
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• v.36 no.2
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• pp.200-208
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• 1993

연두금파리 종령유충의 복강내에 이물질로서 protein-A gold 용액(20 nm 크기) colloidal gold 용액(15 nm 크기), 그리고 이들의 혼합액을 주입하여 혈구가 이들을 처리하는 양상을 전자현미경을 이용하여 관찰하였다. Gold 입자에 대한 혈구의 세포성 면역작용 방식은 식세포작용이었고 이러한 식세포 작용은 연두금파리에서 확인한 prohemocyte, plasmatocyte. granulocyte type I, II, III, 그리고 oneocvtoid 등의 6가지 혈구 중 type 11 granulocyte에 의해서 수행되었다. Gold 입자는 주로 판상의 원형질 돌기를 통하여 혈구 속으로 들어갔으며 coated-pit로 보이는 구조에 의해서도 받아들이는 것으로 나타났다. 식세포 작용은 gold 용액 주입후 10분 이내에 빠른 속도로 일어났으며 시간이 경과함에 따라 gold 입자를 함유한 phagosome은 리소소음과 융합하여 phagolysosome을 형성하면서 세포질내에 축적되어 있었다. Gold 용액을 주입한 후 type II granulocyte에서는 세포질 돌기와 전자 밀도가 높은 과립, 그리고 세포 소기관이 소실되는 등의 변화가 일어났다. protein A-gold 및 colloidal gold 용액에 대한 기본적인 식세포 과정은 별다른 차이점이 없었으나 gold 입자 섭취 초기 과정과 양, 그리고 multivesicular body의 모습등에서는 차이점을 보여 주었다.

• Journal of Pharmaceutical Investigation
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• v.34 no.6
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• pp.465-469
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• 2004

Colloidal gold nanoparticles might be of use as nano scale delivery systems of various therapeutic materials in the future. Recent studies have reported the feasibility of colloidal gold nanoparticles as gene delivery systems or protein delivery systems. In this study, we aimed to develop a short-step method useful for screening the optimal coating conditions of colloidal gold nanoparticles with proteins. We observed that colloidal gold nanoparticles have properties of changing its unique color when they were exposed to NaCl solution. Taking advantage of the color changing properties of colloidal gold nanoparticles, we applied the color testing method of colloidal gold nanoparticles solutions for evaluating the protein coating nature. Using bovine serum albumin as a model protein, we tested the protein coating of colloidal gold nanoparticles via the color change upon NaCl addition. The optimal coating concentration and coating conditions of colloidal gold nanoparticles with bovine serum albumin were fixed using the color testing methods. We suggest that the color testing method might be applied to optimize the coating condition of colloidal gold nanoparticles with other therapeutic proteins.

• Korean Journal of Veterinary Research
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• v.29 no.4
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• pp.541-548
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• 1989

The present study was carried out to determine viral antigens and its morphogenesis in the ultrathin frozen and araldite sections of cell cultures infected with ADV by protein A-gold labeling. ADV antigens were labeled with 10nm gold probes, and electron-dense gold particles were mainly present on viral nucleocapsids and viral envelopes. Immunogold labeling in the ultracryosections showed a very low degree of interaction with tissue structures. Immunogold labeling in the ultrathin cryosections can be useful tool for the detection of ADV antigens, and the technique also may provide its great potential for immunocytochemical studies on various virus-host cell Interactions.

• Research in Plant Disease
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• v.12 no.1
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• pp.25-27
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• 2006

Rice dwarf phytoreovirus (RDV), a member of the family Reoviridae has a genome composed of 12 segmented dsRNAs designated as 51 to 512 with an increasing order of mobility in polyacrylamide gel electrophoresis (PAGE). RDV encode 12 structural and non-structural proteins, $P1{\sim}P12$ which are encoded by the $S1{\sim}S12$ segments of the dsRNA genome, respectively. In this experiment, we confirmed in situ localization of RDV particles and P12 in cytoplasm of infected rice plant. We observed specific reaction of the gold particles using virus particle and P12 protein specific antiserum with protein A-gold immunolabelling in electron microscope. It was observed that gold particles specifically react to virus particles in cytoplasm in case using the antiserum for virus particles. In the case of antiserum for P12 protein, gold particles sporadically existing on cytoplasm without existing in organelle of cytoplasm specifically. As this result, RDV P12 protein encoded by S12 located in cytoplasm.

• Applied Science and Convergence Technology
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• v.24 no.1
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• pp.1-8
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• 2015

Protein immobilization on a gold surface plays an important role in the usefulness of biosensors that utilize gold-coated surfaces such as surface plasmon resonance (SPR), quartz crystal microbalance (QCM), etc. For developing high performance biosensors, it is necessarily required that immobilized proteins must remain biologically active. Loss of protein activity and maintenance of its stability on transducer surfaces is directly associated with the choice of immobilization methods, affecting protein-protein interactions. During the past decade, a variety of strategies have been extensively developed for the effective immobilization of proteins in terms of the orientation, density, and stability of immobilized proteins on analytical devices operating on different principles. In this review, recent advances and novel strategies in protein immobilization technologies developed for biosensors are briefly discussed, thereby providing an useful information for the selection of appropriate immobilization approach.

• Journal of Sensor Science and Technology
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• v.14 no.3
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• pp.137-143
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• 2005

Research in the field of biosensor has enormously increased over the recent years. The metal-oxide semiconductor field effect transistor (MOSFET) type protein sensor offers a lot of potential advantages such as small size and weight, the possibility of automatic packaging at wafer level, on-chip integration of biosensor arrays, and the label-free molecular detection. We fabricated MOSFET protein sensor and proposed the gold-black electrode as the gate metal to improve the response. The experimental results showed that the output voltage of MOSFET protein sensor was varied by concentration of albumin proteins and the gold-black gate increased the response up to maximum 13 % because it has the larger surface area than that of planar-gold gate. It means that the expanded gate allows a larger number of ligands on same area, and makes the more albumin proteins adsorbed on gate receptor.

• Korean Journal of Food Science and Technology
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• v.31 no.1
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• pp.1-6
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• 1999

A piezoelectric (PZ) biosensor system detecting Salmonella spp. was developed. The system consisted of an oscillator, a frequency counter and an antibody-immobilized quartz crystal. An anti-Salmonella antibody was immobilized on one gold. surface of the quartz crystal with protein A. Salmonella detection was made by measuring resonant frequency shift owing to a mass change by specific binding of microbial cells to the gold surface of the PZ crystal. The PZ antibody sensor was operated optimally at 0.1M phosphate buffer, pH 7.2 and $35^{\circ}C$. The sensor was quite specific to Salmonella spp. The obtained frequency shift was correlated with the Salmonella concentration in the range of $10^5{\sim}10^6\;CFU/mL$. The frequency shift increased further by addition of polystyrene beads. The Salmonella detection which is indicated by a steady-state microbial adsorption to the quartz crystal was accomplished within 50min.

• Bulletin of the Korean Chemical Society
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• v.33 no.10
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• pp.3269-3273
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• 2012

In this study, we report a new method for the high contrast imaging of biomolecular interactions at roller printed protein surfaces using thermotropic liquid crystals (LCs). Avidin was roller printed and covalently immobilized onto the obliquely deposited gold surface that was decorated with carboxylic acid-terminated self-assembled monolayers (SAMs). The optical response of LCs on the roller printed film of avidin contrasted sharply with that on the obliquely deposited gold surface. The binding of biotin-peroxidase to the roller printed avidin was then investigated on the obliquely deposited gold substrate. LCs exhibited a non-uniform and random orientation on the roller printed area decorated with the complex of avidin and biotin-peroxidase, while LCs displayed a uniform and planar orientation on the area without roller printed proteins. The orientational transition of LCs from uniform to non-uniform state was triggered by the erasion of nanometer-scale topographies on the roller printed surface after the binding of biotin-peroxidase to the surface-immobilized avidin. The specific binding events of protein-receptor interactions were also confirmed by atomic force microscopy and ellipsometry. These results demonstrate that the roller printing of proteins on obliquely deposited gold substrates could provide a high contrast signal for imaging biomolecular interactions using LC-based sensors.

• Korean Journal Plant Pathology
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• v.14 no.3
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• pp.229-239
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• 1998

Antibodies were raised against fusion proteins of the N-terminus and a region containing the GDNQ (Gly-Asp-Asn-Gln) polymerase motif of the L (polymerase) protein of sonchus yellow net virus (SYNV). Immunoblot analyses using these antibodies revealed the presence of the L protein in purified SYNV preparations and in nuclear extracts from infected tobacco. The serological analyses and detection in a polyacrylamide gels suggested that the L protein is present in at least a 20 fold lower abundance than the G, N, M1 and M2 proteins, and has size corresponding to a molecular weight of over 200 kDa as predicted from nucleotide sequence data. Electron microscopy with gold-labelled antibodies was used to localize the N, M2, and G proteins of SYNV in thin sections of infected tissue. When sections of SYNV-infected tissue were treated with antisera against total SYNV proteins and N protein, gold label could be detected in both the viroplasms and in virus particles. With the anti-M2 protein antiserum, the gold label was strongly localized in the viroplasms but only limited labelling of the virus particle sonly. Limited labelling of the L protein was observed in the viroplasms and the virus particles, presumably because of the low abundance of L protein in the tissues.

• Bulletin of the Korean Chemical Society
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• v.34 no.9
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• pp.2725-2730
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• 2013

Single C-reactive protein (CRP) molecules, which are non-specific acute phase markers and products of the innate immune system, were quantitatively detected on a gold-nanopatterned biochip using evanescent field-enhanced fluorescence imaging. The $4{\times}5$ gold-nanopatterned biochip (spot diameter of 500 nm) was fabricated by electron beam nanolithography. Unlabeled CRP molecules in human serum were identified with single-molecule sandwich immunoassay by detecting secondary fluorescence generated by total internal reflection fluorescence (TIRF) microscopy. With decreased standard CRP concentrations, relative fluorescence intensities reduced in the range of 33.3 zM-800 pM. To enhance fluorescence intensities in TIRF images, the distance between biochip surface and CRP molecules was optimally adjusted by considering the quenching effect of gold and the evanescent field intensity. As a result, TIRF only detected one single-CRP molecule on the biochip the first time.