• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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• pp.84-84
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• 2013

FcBP consisting of 13 amino acids specifically binds to Immunoglobulin G Fc domain. Initially, we utilized this peptide for preparation of antibody chip as a PEG composite for enhanced solubility. After then, the peptide conjugate was immobilized on agarose resin, resulting in highly efficient affinity column for antibody purification. The efficiency was comparable to commercial Protein A column. Recently, this peptide was conjugated with cell penetratingpeptide (CPP) on a backbone of GFP, affording antibody transducer, which carries antibody into live cells by simple mixing of antibody and the transducer in cell culture media. Antibody transduction into cells was monitored by live cell imaging. More recently, the FcBP was fused to ferritin cage, which consists of 24 ferritin protein molecules. The FcBP-ferritin cage showed greatly increased binding affinity to human IgG. Its binding was analyzed by QCM and SPR analysis. Finally, it was selectively delivered by Herceptin to SKBR3, a breast cancer cell, over MCF10A, non-tumorigenic cells.

• Journal of Microbiology and Biotechnology
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• 제17권6호
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• pp.1036-1040
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• 2007

Sodium butyrate (NaBu) has been used to enhance protein expression levels in mammalian cell culture. To determine the clonal variability of recombinant Chinese hamster ovary (rCHO) cells in response to NaBu addition regarding specific antibody productivity $(q_{Ab})$, three rCHO clones were subjected to different concentrations of NaBu. For all three clones, NaBu addition inhibited cell growth and decreased cell viability in a dose-dependent manner. On the other hand, the enhancing effect of NaBu on $q_{Ab}$ varied significantly among the clones. NaBu addition enhanced the antibody production of only one clone. RT-PCR analysis revealed that the changes in $q_{Ab}$ correlated linearly with those of the mRNA transcription level. Thus, it was concluded that the different enhancing effects of NaBu on protein expression in rCHO cell clones resulted from their different mRNA transcription levels.

• 한국발생생물학회지:발생과생식
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• 제28권3호
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• pp.75-86
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• 2024

Artificial sexual maturation of eel (Anguilla japonica) involves rearing in seawater and injecting salmon pituitary extract (SPE). The salinity of seawater and components of SPE influence hormonal activities of the eel pituitary, leading to gonad development. This study investigated the direct effects of salinity change and SPE treatment on the eel pituitary gland using primary cell cultures. Pituitary cells were cultured into four experimental groups: control culture (control), SPE-treated culture (SPE), NaCl-treated culture (NaCl) and NaCl+SPE NaCl+SPE treated culture (NaCl+SPE). We investigated the expression of genes presumably related to reproduction and/or salinity, including luteinizing hormone (LHβ), follicle stimulating hormone (FSHβ), progesterone receptor-like (pgrl), prolactin (PRL), dopamine receptor D4 (drd4), neuropeptide B/W receptor 2 (NPBWR2) and relaxin family peptide receptor 3-2b (rxfp3-2b). Gene expression analysis revealed significant upregulation of LHβ in SPE and NaCl+SPENaCl+SPE groups compared to control and NaCl (p<0.05). FSHβ expression did not show any significant changes. PRL showed a significant decrease in the NaCl group (p<0.05). Pgrl, NPBWR2, drd4, and rxfp3-2b displayed the highest expression in the control group, with downregulation observed in all treatment groups (NaCl, SPE, and NaCl+SPE) (p<0.05). This study demonstrated the direct effects of salinity changes and SPE treatment on the eel pituitary. Results from this study also suggest that salinity change is necessary but work together with SPE to induce reproductive process, and that LHβ, pgrl, PRL, drd4, NPBWR2, and rxfp3-2b genes are obviously associated with reproduction and salinity changes in eels.

• 대한한방부인과학회지
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• 제22권1호
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• pp.125-139
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• 2009

Purpose: This experiment was designed to find out increasing effects of S. barbata. co-treatment with anti-cancer drugs at cancer cell's growth inhibition effect. Methods: Divergent observational study of the S. barbata. co-treatment with Cisplatin treatment on HeLa cell. Cell viability using MTT assay, Cell Culture and Cytotoxicity Studies, Cell Cycle Analysis, Annexin V-FITC/PI assay, Cell morphological assessment, PARP cleavage using Western blotting analysis when HeLa cell were co-treated with Cisplatin and Scutellaria Barbata extracts. Results: When HeLa cell were co-treated with Cisplatin and Scutellaria Barbata extracts, we found out viability of HeLa cell, changing in the distribution of cell cycle, Annexin V-FITC staining, DAPI staining, PARP clavage protein assay by Western-blot. So Scutellaria Barbata extracts have increased apoptosis Conclusion: When co-treated Scutellaria Barbata extracts with anti-cancer drugs, the anti-cancer effects were increased. We still not sure which constituent apoptosis at cancer cells and activates anti-cancer effects suppressing, but we believe that it'll be revealed here after with following experiments.

• 대한한방소아과학회지
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• 제30권4호
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• pp.29-59
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• 2016

Objectives PRAL (Prunus armeniaca Linne Var) has been known to suppress allergic reaction. However, the cellular target and its mechanism of action were unclear. This study was designed to investigate the effect of PRAL on RBL-2H3 mast cell, which is PMA-Ionomycin-induced activated in vitro and the effect of PRAL on the MNC/Nga mice that are DNCB-induced activated in vivo. Methods In this study, IL-4, IL-13 production were examined by ELISA analysis; IL-4, IL-13, IL-31, IL-31Ra, $TNF-{\alpha}$ and GM-CSF mRNA expression were examined by Real-time PCR; manifestations of AP-1 and MAPKs transcription factors were examined by western blotting in vitro. Then skin rashes have been evaluated and verified the distribution of mast cells by H&E and toluidine blue. Also, WBC, eosinophil and neutrophil, IgE level in serum, $IFN-{\gamma}$, IL-4, IL-5 in the splenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$, $CD3^+CD69^+$ in the Axillary Lymph Node (ALN), PBMCs and dorsal skin and IL-5, IL-13, IL-31, IL-31Ra in the dorsal skin by Real-time PCR were all evaluated from the NC/BNga mice. Results As a result of this study, the mRNA expression of IL-4, IL-13, IL-31, IL-31Ra and $TNF-{\alpha}$ and IL-4, IL-13 production, shown in ELISA analysis, were suppressed by PRAL. Results from the western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including AP-1 and p-JNK, p-ERK. Histological examination showed that infiltration levels of immune cells in the skin of the AD-induced NC/Nga mice were improved by PRAL orally adminstration. Orally- administered PRAL group also showred decreased level of IgE in the serum. This group has shown decreased the level of IL-4, IL-5, but shown elevated $IFN-{\gamma}$ level in the splenocyte culture supernatant. The same group also has shown decreased cell numbers of $CD4^+$, $CD8^+$, $CD3^+CD69^+$ in the ALN, and $CD4^+$, $Gr-1^+CD11b^+$ in the dorsal skin. PRAL oral adminstration increased cell numbers of $CD4^+$, but decreased cell numbers of $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$ in the PBMCs. Conclusions Obtained results suggest that PRAL can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis (AD) induced in the NC/Nga mice. This may play an important role in recovering AD symptoms and suppressing pruritus.

• Journal of Periodontal and Implant Science
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• 제41권1호
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• pp.17-22
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• 2011

Purpose: Nitric oxide (NO) has been known as an important regulator of osteoblasts and periodontal ligament cell activity. This study was performed to investigate the relationship between NO-mediated cell death of human periodontal ligament fibroblasts (PDLFs) and N-methyl-D-aspartic acid (NMDA) receptor antagonist (+)-5-methyl-10, 11-dihydro-5H-dibenzo[a,d]cyclohepten-5, 10-imine hydrogen maleate (MK801). Methods: Human PDLFs were treated with various concentrations (0 to 4 mM) of sodium nitroprusside (SNP) with or without $200\;{\mu}M$ MK801 in culture media for 16 hours and the cell medium was then removed and replaced by fresh medium containing MTS reagent for cell proliferation assay. Western blot analysis was performed to investigate the effects of SNP on the expression of Bax, cytochrome c, and caspase-3 proteins. The differences for each value among the sample groups were compared using analysis of variance with 95% confidence intervals. Results: In the case of SNP treatment, as a NO donor, cell viability was significantly decreased in a concentration-dependent manner. In addition, a synergistic effect was shown when both SNP and NMDA receptor antagonist was added to the medium. SNP treated PDLFs exhibited a round shape in culture conditions and were dramatically reduced in cell number. SNP treatment also increased levels of apoptotic marker protein, such as Bax and cytochrome c, and reduced caspase-3 in PDLFs. Mitogen-activated protein kinase signaling was activated by treatment of SNP and NMDA receptor antagonist. Conclusions: These results suggest that excessive production of NO may induce apoptosis and that NMDA receptor may modulate NO-induced apoptosis in PDLFs.

• 한국해양바이오학회지
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• 제15권2호
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• pp.82-89
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• 2023

Microalgae, as photosynthetic organisms, possess the ability to produce a diverse array of bioactive compounds. This study focused on the transformant Chlamydomonas reinhardtii dZL and subjected it to cultivation under varying light intensities (60, 120, 180, and 240 µmol/m²/s). Our aim was to assess the impact of light intensity on both microalgal biomass and carotenoid production. The cultivation took place in 80 mL bubble column photobioreactors, specifically the Multi-cultivator. Notably, the culture exposed to 240 µmol/m²/s exhibited the most rapid cell growth, surpassing even the cell concentration achieved at 180 µmol/m²/s by day 8. A detailed analysis of the specific irradiance rate over time unequivocally revealed a sharp decline in growth rates when the rate fell below 2 × 10^-10 µmol/cell/s. Although the culture with 60 µmol/m²/s yielded the highest carotenoid content (1.2% of dry weight), the culture exposed to 240 µmol/m²/s recorded the highest carotenoid concentration at 8.9 mg/L owing to its higher biomass. Our findings reveal the critical importance of maintaining a specific irradiance rate above 2 × 10^-10 µmol/cell/s to enhance biomass and carotenoid productivity. This study lays the groundwork for defining optimal light intensity conditions applicable to mass culture systems, with the objective of augmenting C. reinhardtii biomass and optimizing carotenoid productivity.

• Journal of Microbiology and Biotechnology
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• 제12권5호
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• pp.793-800
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• 2002

The mycolic acids and fatty acids of mycolic acid- containing bacteria in various types of fluids were analyzed using capillary gas chromatography and mass spectrometry. As model strains, Brevibacterium and Coryebacterium species, which have corynomycolic acids ill the range of $C_{32}C_{36}$ in the whole cell, were investigated. Optimized solvents extraction procedures for the mycolic acids and fatty acids from the culture fluids were: chloroform/methanol (1:2, v/v) as the first extraction solvents fur 4 h; and chlorofunuwater (1:1, v/v) as the second extraction solvents far 1 h. These conditions gave above 95% recovery yields fur mycolic acids from the culture fluids. The mycolic acid profile for the whole cells and the culture fluids were similar fur all the media tested. Thus, the procedure described here could be applied for the identification of mycolic acid-containing bacteria in fermentation broth or liquid from of foods.

• Journal of Applied Biological Chemistry
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• 제50권4호
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• pp.221-226
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• 2007

The high molecular-weight mucilage extracted and purified from cactus fruit of Opuntia ficus-indica var. Saboten was degraded by the cell-free culture filtrate of a fungus isolated from soil. TLC analysis of the polymeric mucilage after incubation with the fungal culture filtrate confirmed its degradation. When the degradation products were tested for their qualitative reactions with ninhydrin and phenol-sulfuric acid, only phenol-sulfuric acid gave positive development, and ninhydrin did not show any observable color reaction. This coloring reaction suggested the presence of a carbohydrate without an amino group within the mucilage. Analyses by HPLC and liquid gel permeation chromatography on sephadex G-100 also provided additional information on degradation of the mucilage by the fungal culture filtrate. The sequences of ITS-5.8S rDNA from the fungal isolate that was cultivated for the preparation of mucilage-degrading enzyme showed 99% similarity to those of Pestalotiopsis aquatica.

• 대한수의학회지
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• 제26권2호
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• pp.245-249
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• 1986

The present study has been carried out to investigate the optimal condition on the blastogenesis of guinea pig lymphocytes. A microculture system in conjunction with a semiautomatic multiple sample harvester(SAMSH) was used to study the in vito optimal condition of guinea pig lymphocytes. Data were presented to show many variables that are Involved in studying the responses of guinea pig lymphocyte in a microculture system to the stimulation of Concanavalin A(Con A) and lipopolysaccharide (LPS). Analysis indicated that the conditions for optimal Con A as measured by incorporation of $^3H$-TdR include : (1) use of RPMI-1640 as culture medium, (2) use of $6{\mu}g$ of Con A, per culture, (3) use of $1{\times}10^6$ cells per culture. Conditions for optimal stimulation with LPS mitogen were similar to those used for Con A.