• Proceedings of the Korea Society of Poultry Science Conference
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• 2004.11a
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• pp.9-10
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• 2004

We developed a new panel of markers for the characterization of chicken PGCs. The results of immunostaining demonstrated that anti-SSEA-3, anti-SSEA-4, anti-integrin 6, and anti-integrin 1 antibodies. and STA and DBA bound specifically to chicken PGCs. These reagents could be used to characterize chicken PGCs together with conventional marker reagents such as PAS and anti-SSEA-1 antibody. We also showed that double staining of PGCs with the newly developed markers was feasible, which might contribute to rapid detection and accurate characterization of chicken PGCs.

• BMB Reports
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• v.33 no.2
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• pp.97-102
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• 2000

Phosphoinositide-specific phospholipase C-${\gamma}1$ (PLC-${\gamma}1$) is a pivotal mediator in the signal transduction cascades induced by many growth factors. Using a yeast two-hybrid system, heat-shock protein 90 (Hsp90) was identified as a PLC-${\gamma}1$-binding protein. A co-immunoprecipitation experiment, using anti-PLC-${\gamma}1$ antibody, demonstrated an in vivo interaction between Hsp90 and PLC-${\gamma}1$ in the NIH-3T3 cells. The interaction in NIH-3T3 was unaffected by the PDGF treatment, inducing phosphorylation and activation of PLC-${\gamma}1$. Direct interaction between Hsp90 and PLC-${\gamma}1$ was confirmed by in vitro binding experiments using purified Hsp90 and PLC-${\gamma}1$. Furthermore, Hsp90 increased the $PIP_2$-hydrolyzing activity of PLC-${\gamma}1$ up to 2-fold at $0.1{\mu}M$ in vitro. Taken together, we show for the first time, the interaction of PLC-${\gamma}1$ with Hsp90, both in vivo and in vitro. We suggest that Hsp90 may play a role in PLC-${\gamma}1$-mediated signal transduction.

• Journal of Microbiology and Biotechnology
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• v.16 no.12
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• pp.1968-1976
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• 2006

Recombinant E. coli ACV 1003 (recA:: lacZ) was used to measure low concentrations of DNA-damaging chemicals, which produce $\beta$-galactosidase via an SOS regulon system. Very low $\beta$-galactosidase activities of less than 0.01 unit/ml, $\beta$-galactosidase produced through an SOS response corresponding to the 10 ng/ml (ppb) of DNA damaging chemicals in the environment, can be rapidly determined by using an alternative interferometric biosensor with optically flat thin films of porous silicon rather than by the conventional time-consuming Miller's enzyme assay as well as the ELISA method. fu order to enhance the sensitivity in the interferometry, it needs to obtain more uniform distribution and higher biolinking efficiency, whereas interferometric sensing is rapid, cheap, and advantageous in high throughput by using a multiple-well-type chip. In this study, pore size adjusted to 60 nm for the target enzyme $\beta$-galactosidase to be bound on both walls of a Si pore and a calyx crown derivative was apllied as a more efficient biolinker. Furthermore, anti-$\beta$-galactosidase was previously functionalized with the biolinker for the target $\beta$-galactosidase to be specifically bound. When anti-$\beta$-galactosidase was bound to the calyx-crown derivative-linked surface, the effective optical thickness was found to be three times as high as that obtained without using anti-$\beta$-galactosidase. The resolution obtained was very similar to that afforded by the time-consuming ELISA method; however, the reproducibility was still unsatisfactory, below 1 unit $\beta$-galactosidase/ml, owing to the microscopic non-uniform distribution of the pores in the etched silicon surface.

• Journal of the Korean Society of Manufacturing Process Engineers
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• v.15 no.6
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• pp.122-128
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• 2016

The Biosensor biosensor pattern was developed by via an EHD (electro-hydro-dynamics (EHD) patterning process that was performed under atmospheric pressure at room temperature in a single step. The drop diameter was smaller than nozzle diameter and applied high viscosity conductive ink was applied in the EHD patterning method to provide a clear advantage over the piezo and thermal inkjet printing techniques. The Biosensor's biosensor's micro electrode pattern was printed by via a continuous EHD patterning method using 3three- type types of control parameters parameter (input voltage, patterning speed, nozzle pressure). High viscosity (1000 cps) conductive ink with 75 wt% of silver nanoparticles was used for experimentation. The incremental result of impedance of biosensor impedance was measured between the antibody ($10ug{\mu}g/ml$) to spore (0.1 ng/ml, 10 ng/ml, and $1ug{\mu}g./ml$) reaction at frequency 493 MHz frequency.

• Korean Journal of Agricultural Science
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• v.39 no.4
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• pp.613-618
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• 2012

This study was performed to optimize the basic platform of a lateral flow immunoassay. Improvement of the limit of detection (LOD) was evaluated according to the width of a nitrocellulose membrane with varying concentrations of analyte. The analyte, neutravidin was detected based on the avidin-biotin interaction. The antibody-Au nanoparticle conjugation was mostly stabled in a PBS buffer of pH 7.3. The optimal widths of a nitrocellulose membrane were 4 and 6 mm considering the sample flow rate and signal strength of the test line on the membrane. The LOD of neutravidin was 0.001 mg/ml in the optimum conditions.

• BMB Reports
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• v.33 no.6
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• pp.483-492
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• 2000

The Herpes simplex virus type 1 (HSV-1) strain F glycoprotein H (gH) gene in the pHLB-4 plasmid was recombinated into a baculovirus expression vector (lacZ-HcNPV) to construct a recombinant virus GH-HcNPV expressing gH. The sequences of gH and its expression were analyzed. The gH gene was located in the 6.41 kb BglII fragment. The open reading frame (ORF) of the gH gene was 2,517 bp and codes 838 amino acid residues. Insect cells infected with this recombinant virus synthesized a high level of the matured and gX-gH fusion protein with approximately 112 kDa. The fusion gH protein was localized on the membrane of the insect cells as seen by using immunofluorescence assay and accumulated in the cultured media by the SDS-PAGE and immunoprecipitation assays. The amino acid sequence presents additional characteristics compatible with the structure of a viral glycoprotein: signal peptide, putative glycosylation sites and a long C-terminal transmembrane sequence. Antibodies raised in mice to this recombinant protein recognized viral gH and neutralized the infectivity of HSV-1 in vitro. These results demonstrate that it is possible to produce a mature protein by gene transfer in eukaryotic cells, and indicate the utility of the HcNPV-insect cell system for producing and characterizing eukaryotic proteins. Furthermore, the neutralizing antibodies would appear to protect mice against HSV; accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

• YAKHAK HOEJI
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• v.38 no.1
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• pp.6-11
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• 1994

The MeOH extract of the fruits of Melia toosendan was selected for futher study by its cytotoxicity and effect on the human breast cancer cell line, MCF-7. The active principle obtained by activity guided fractionation followed by purification gave rise to a needle crystal. The structure was deduced by employing NMR and was determined to be identical with 28-deacetyl sendanin by comparison with published data. This compound induced morphological change of MCF-7 to be rounded with tubule at concentrations between $50\;{\mu}g/ml$ and $0.025\;{\mu}g/ml$. This compound, however, showed strong cytotoxic effect on Hepalclc7 and HepG2, and their $GI_{50}$ on the hepatoma cell lines were $0.238\;{\mu}g/ml$ and $0.805\;{\mu}g/ml$, respectively. Its effect on lymphocyte of mouse was stronger than hepatoma cell lines, and their $ED_{50}$ of polyclonal antibody response was $0.011\;{\mu}g/ml$, and $ED_{50}$ of cell viability was $0.039\;{\mu}g/ml$.

• BMB Reports
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• v.40 no.6
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• pp.1028-1038
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• 2007

Human plasma Hp is classified as 1-1, 2-1, and 2-2. They are inherited from two alleles Hp 1 and Hp 2, but there is only Hp 1 in almost all the animal species. Hp 2-2 molecule is extremely large and heterogeneous associated with the development of inflammatory-related diseases. In this study, we expressed entire bovine Hp in E. coli as a $\alpha\beta$ linear form. Interestingly, the antibodies prepared against this form could recognize the subunit of native Hp. In stead of a complicated column method, the antibody was able to isolate bovine Hp via immunoaffinity and gelfiltration columns. The isolated Hp is polymeric containing two major molecular forms (660 and 730 kDa). Their size and hemoglobin binding complex are significantly larger than that of human Hp 2-2. The amino-acid sequence deducted from the nucleotide sequence is similar to human Hp 2 containing a tandem repeat over the $\alpha$ chain. Thus, the Hp 2 allele is not unique in human. We also found that there is one additional -SH group (Cys-97) in bovine $\alpha$ chain with a total of 8 -SH groups, which may be responsible for the overall polymeric structure that is markedly different from human Hp 2-2. The significance of the finding and its relationship to structural evolution are also discussed.

• Applied Chemistry for Engineering
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• v.33 no.2
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• pp.173-178
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• 2022

A lateral flow immunoassay (LFIA) method using carbon nanodot@silica as a signaling material was developed for analyzing the concentration of retinol-binding protein 4 (RBP4), one of the lung cancer biomarkers. Instead of antibodies mainly used as bioreceptors in nitrocellulose membranes in LFIA for protein detection, aptamers that are more economical, easy to store for a long time, and have strong affinities toward specific target proteins were used. A 5' terminal of biotin-modified aptamer specific to RBP4 was first reacted with neutravidin followed by spraying the mixture on the membrane in order to immobilize the aptamer in a porous membrane by the strong binding affinity between biotin and neutravidin. Carbon nanodot@silica nanoparticles with blue fluorescent signal covalently conjugated to the RBP4 antibody, and RBP4 were injected in a lateral flow manner on to the surface bound aptamer to form a sandwich complex. Surfactant concentrations, ionic strength, and additional blocking reagents were added to the running buffer solution to optimize the fluorescent signal off from the sandwich complex which was correlated to the concentration of RBP4. A 10 mM Tris (pH 7.4) running buffer containing 150 mM NaCl and 0.05% Tween-20 with 0.6 M ethanolamine as a blocking agent showed the optimum assay condition for carbon nanodot@silica-based LFIA. The results indicate that an aptamer, more economical and easier to store for a long time can be used as an alternative immobilizing probe for antibody in a LFIA device which can be used as a point-of-care diagnosis kit for lung cancer diseases.

• Applied Chemistry for Engineering
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• v.25 no.5
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• pp.544-547
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• 2014

In this study, an enzyme-linked immunosorbent assay (ELISA) and immuno-chromatographic technique were combined to fabricate immuno-strip sensors for the detection of Legionella pneumophila. The immuno-strip sensor was manufactured with four different membranes. A nitrocellulose membrane was used to immobilize capture antibody and generate signals due to the high affinity to antibodies, and glass fiber membranes were used as a conjugate release pad and a sample application pad. A cellulose membrane was used as an absorption pad to induce sample flow by the capillarity. Colorimetric signals produced by sandwich immuno-reaction and enzyme reaction could be analyzed qualitatively and quantitatively within 30 min. Under the given experimental conditions, sensor signals with L. pneumophila samples were observed qualitatively by naked eyes and measured quantitatively in a range of $1.3{\times}10^3-1.3{\times}10^6CFU/mL$ with a digital camera and home-made image analysis software.