• KSBB Journal
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• v.21 no.2
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• pp.133-139
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• 2006

In 'solid-phase' PEGylation, the conjugation reaction occurs as the proteins are attached to a solid matrix, and thus it can have distinct advantages over the conventional, solution-phase process. We report a case study: rhIFN-${\alpha}$-2a was first adsorbed to cation exchange resin and then N-terminally PEGylated by aldehyde mPEG of 5, 10, and 20 kD through reductive alkylation. After the PEGylation, salt gradient elution efficiently recovered the mono-PEGylate in a purified form from the unwanted species such as unmodified IFN, unreacted PEG, and others. The mono-PEGylation and its purification were integrated in a single chromatographic step. Depending on the molecular weight of the mPEG aldehyde used, the mono-PEGylation yield ranged 50-64%. We could overcome the major problems of random, or uncontrollable, multi-PEGylation and the post-PEGylation purification difficulties associated with the solution-phase process. N-terminal sequencing and MALDI-TOF MS confirmed that a PEG molecule was conjugated only to the N-terminus. Compared with the unmodified IFN, the mono-PEGylate showed the reduced anti-viral activity as measured by the cell proliferation assay. The bioactivity was reduced more as the higher molecular weight PEG was conjugated. Immunoreactivity, evaluated indirectly by antibody binding activity using a surface plasmon resonance biosensor, also decreased. Nevertheless, trypsin resistance as well as thermal stability was considerably improved.

• Journal of Biomedical Engineering Research
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• v.15 no.1
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• pp.41-50
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• 1994

The chemotherapy using macromolecules, i.e., monoclonal antibodies loaded with anticancer agents hasn't been successful in delivering therapeutic amount of the conjugates. The comprehensive evaluation of mass transfer factors in tumor is prerequisite for the development of the effective chemotherapy. Characterization of neuroblastoma implanted in immunosuppressed athymic nude rats was performed. Its growth kinetics, glucose metabolic rate (GMR) were measured along with the interstitial fluid pressure (IFP), blood perfusion rate (BPR) and pH distribution throughtout the tumor radius. Volume doubling time and GMR were 8.1 days(SD 0.44 day), 23.53 mg/min/100 g(SD 3.54 mg/min/100 g), respectively. The IFP in tumor center was increased with tumor volume, and approached to 3 mmHg (SD 2.6 mmHg) when the tumor was 3 cm high. The radial distribution of IFP, BPR and pH in 2 cm high tumors showed that BPR and pH were decreased, while IFP was increased as the ~ensors moved toward the tumor center. The elevated IFP, decreased BPR and pH in tumor center suggested that the delivery of conjugates might be increased by properly manipulating mass transfer factors.

• Journal of the Korean Applied Science and Technology
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• v.41 no.2
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• pp.159-171
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• 2024

This study confirmed the antioxidant activity and antimicrobial efficacy and formulation stability for the effectiveness experiment of Rumex crispus. L root extract. For antioxidant activity, DPPH radical scavenging, FRAP activity, ABTS+ radical scavenging, and SOD-like activity were performed. Antimicrobial activity was evaluated for Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Candida albicans strains. In addition, skin containing Rumex crispus. L root extract is checked over time for pH, temperature, and daylight for 21 days. As a result of antioxidant evaluation, it was confirmed that the activity increased in a concentration-dependent manner at a concentration of 0.0625-1 mg/mL. The clear zones of each bacterium at 100mg/mL concentrations were 10.45±0.34, 9.77±0.59, 9.92±0.22, and 10.08±0.12, which were superior to the control group Methyl paraben, and the antibacterial power of S. aureus and E. coli was confirmed at 100mg/mL concentration for MIC. There was little change in absorbance when the pH of the skin was 4.0, 6.0, and 7.0 and At 4℃, 25℃, and 40℃, it was discolored as the temperature increased. It was also observed that discoloration occurred when exposed to daylight. This is presumed to be able to prevent discoloration when it is shielded and stored at low temperatures. When the results of this study are summarized, Rumex crispus. L root extract is considered to have high value in use as a cosmetic raw material that can expect antioxidant and antibacterial activities.

• Korean Chemical Engineering Research
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• v.44 no.1
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• pp.65-72
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• 2006

Recent technical advances in the biorecognition engineering and the microparticle fabrication may enable us to develop the single step purification using magnetic particle, because of its simplicity, efficacy, ease of automation, and process economics. In this study, we used commercial magnetic particles from Seradyn, Inc. (Indianapolis, USA). It was ca. 2.8 micron in diameter, consisted of polystyrene core and magnetite coating, and its surface had carboxyl groups. The model, capture protein was IgG and anti-IgG was used as the ligand molecule. We studied the different surfaces ('nude', ester-activated, and anti-IgG coated) for their biorecognition of IgG. At a high pH condition, we could reduce non-specific binding. Also anti-IgG immobilized magnetic particle could capture IgG more selectively. We attempted 'oriented immobilization' of anti-IgG, in which the polysaccharides moiety near the C-terminus was selectively oxidized and linked to the hydrazine-coated MP, to improve the efficacy of biorecognitive binding. Using this method, the IgG capturing ability was improved by ca. 2 fold. From the binary mixture of the IgG-insulin, IgG could be more selectively captured. In summary, the oriented immobilization of oxidized anti-IgG proved to be as effective as the streptavidin-biotin system and yet simpler and cost-effective. This immobilization method can find its applications in protein biochips and biotargeting.

• Journal of Life Science
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• v.32 no.12
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• pp.919-928
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• 2022

As seen in the COVID-19 pandemic, unexpected emergence of new viruses presents serious concern on public health. Especially, the absence of effective vaccines or antiviral drugs against emerging viruses significantly increases the severity of disease and duration of viral circulation among population. Natural products have served as a major source for safe and effective antiviral drugs. In this study, we examined the virucidal activity of medical herb extracts with a view to discover novel antiviral agents with desired levels of safety and antiviral efficacy. Ethanol extracts of ten selected medical herbs were tested for antioxidant activity and in-vitro cytotoxicity in various animal cell lines. Of note, the herbal extracts showed broad and potent virucidal activities against rotavirus, hepatitis A virus, and influenza A virus. The extracts of Sorbus commixta and Glycyrrhiza uralensis showed strong virucidal activities against influenza A virus. We also examined whether the extracts of Sorbus commixta and Glycyrrhiza uralensis can be used as inactivating agents to prepare an inactivated viral vaccine. In a mouse model, influenza A virus inactivated by the extracts elicited high levels of neutralizing antibodies, and the vaccination provided complete protection against lethal challenge. These results suggest that herb-derived natural products can be developed to antiviral drugs as well as inactivating agents for preparation of inactivated viral vaccines.

• The Journal of the Korean bone and joint tumor society
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• v.15 no.2
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• pp.130-137
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• 2009

Purpose: STAT3 is an oncogene that regulates critical cellular processes, and its constitutive activation has been demonstrated to correlate with biological and clinical features in many types of human malignancy. Materials and Methods: In this study, STAT3 activation was assessed in variable benign and malignant chondroid tumors in bone by immunohistochemistry using a monoclonal antibody specific for $tyrosine^{705}$-phosphorylated STAT3 ($pSTAT3^{tyr705}$). Results: Among conventional chondrosarcomas (n=17), three cases(50%) of grade III chondrosarcomas were pSTAT3-positive. All grade I and II chondrosarcomas were pSTAT3-negative. This pSTAT3 positivity according to the histologic grade was statistically significant (p=0.0432). Two cases(50%) of clear cell chondrosarcomas were pSTAT3-positive. Six cases (50%) among 12 benign chondroid tumors(6 enchondromas, 3 chondroblastomas, and 3 chondromyxoid fibromas) were also $pSTAT3^{tyr705}$-positive. Conclusion: In conclusion, STAT3 activation is associated with higher tumor grade in conventional chondrosarcomas. Our results suggest that STAT3 is activated in a subset of benign and malignant chondroid tumors, and may support the extension of the cancer stem cell hypothesis to include tumors of cartilaginous lineage.

• Journal of Microbiology and Biotechnology
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• v.18 no.2
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• pp.263-269
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• 2008

Hydrazinocurcumin (HC), a synthetic derivative of curcumin, has been reported to inhibit angiogenesis via unknown mechanisms. Understanding the molecular mechanisms of the drug's action is important for the development of improved compounds with better pharmacological properties. A genome-wide drug-induced haploinsufficiency screening of fission yeast gene deletion mutants has been applied to identify drug targets of HC. As a first step, the 50% inhibition concentration $(IC_{50})$ of HC was determined to be $2.2{\mu}M$. The initial screening of 4,158 mutants in 384-well plates using robotics was performed at concentrations of 2, 3, and $4{\mu}M$. A second screening was performed to detect sensitivity to HC on the plates. The first screening revealed 178 candidates, and the second screening resulted in 13 candidates, following the elimination of 165 false positives. Final filtering of the condition-dependent haploinsufficient genes gave eight target genes. Analysis of the specific targets of HC has shown that they are related to septum formation and the general transcription processes, which may be related to histone acetyltransferase. The target mutants showed 65% growth inhibition in response to HC compared with wild-type controls, as shown by liquid culture assay.

• Journal of Life Science
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• v.30 no.5
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• pp.420-427
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• 2020

A repebody, an artificial non-immunoglobulin protein scaffold, is expected to be a solution in the search for faster, cheaper, and customizable antibodies. However, the production of medical repebodies remains difficult due to their low yield and the complex purification processes required. The Pseudomonas fluorescens ABC transporter system has been suggested as an efficient and cost-effective method for repebody production, but the total yield is low because of the secreted protein's positive charge; thus, a repebody with a high isoelectric point needs to be changed into a more negatively charged protein for better secretion. To achieve this, we first attached oligo-aspartic acids to the N- and C-terminals of the repebody, but secretion efficiency was not enhanced significantly. Subsequently, we devised an alternative method for improved secretion efficiency by engineering fifteen positively charged amino acids to aspartic acid in the non-antigen binding sites of the repebody to give a high net negative charge. As a result, secretion efficiency was greatly enhanced from 21.2% (wildtype) to 58.5% (negatively supercharged). The negatively supercharged repebody was succussfully produced extracellularly by ABC transporter secretion system in P. fluorescens.

• KSBB Journal
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• v.24 no.1
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• pp.1-8
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• 2009

In the past decade, we have observed rapid advances in the development of biochips in many fields including medical and environmental monitoring. Biochip experiments involve immobilizing a ligand on a solid substrate surface, and monitoring its interaction with an analyte in a sample solution. Metal nanoparticles can display extinction bands on their surfaces. These charge density oscillations are simply known as the localized surface plasmon resonance (LSPR). The high sensitivity of LSPR has been utilized to design biochips for the label-free detection of biomolecular interactions with various ligands. LSPR-based optical biochips and biosensors are easy to fabricate, and the apparatus cost for the evaluation of optical characteristics is lower than that for the conventional surface plasmon resonance apparatus. Furthermore, the operation procedure has become more convenient as it does not require labeling procedure. In this paper, we review the recent advances in LSPR research and also describe the LSPR-based optical biosensor constructed with a core-shell dielectric nanoparticle biochip for its application to label-free biomolecular detections such as antigen-antibody interaction.

• Journal of The Korean Association For Science Education
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• v.18 no.4
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• pp.463-472
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• 1998

Biotechnology is the process of using biological system for the production of materials. Genetic engineering, a subset of biotechnology, is the process of altering biological systems by the purposeful manipulation of DNA It is a new field in biology and no topic in biology is more likely to impact our personal lives and is therefore more worthy of our attention and understanding. The purpose of this study was to investigate students' understanding of the concepts of biotechnology, and a test tool which is made up of 20 basic questions was developed for the study. The subject of this study was high school students and the sample size was 486. In order to find out the source of students' misunderstanding, we also analysed high school textbooks and teachers were given the same tool applied to students. Two-way ANOVA was used for the analysis. Major findings of this study are as following; 1. Mean score of students was 41, and there was a significant difference between the scores of boys and girls(p<0.05). Female students scored higher than male students. The variables "region" and "major" had no significant influence. 2. Students' the most misunderstood concepts were "monoclonal antibody" and "gene cloning". Many students thought that a plamid DNA originally has a useful DNA in it, which is apparently wrong. 3. Mean score of teachers was 82, and the variabes of gender and career did not have statistically significant influence on the result(p>0.05). 4. Teachers got the lowest scores on the concepts of "gene therapy", "the accomplishment of biotechnology in agriculture and medicine", and "plasmid DNA". The results of item analysis implied that teachers' misunderstanding might be a part of the sources of students' misunderstaning. 5. Out of 18 basic concepts selected in the study, only 10 concepts were explained well enough in most textbooks. The results of item analysis indicated that textbooks also could be a part of the source of students' misunderstanding.