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Seongsu Kang,Bolam Kim,Se-Jun Yim,Jin-Oh Kim,Dong-Pyo Kim,Yeu-Chun Kim 한국공업화학회 2020 Journal of Industrial and Engineering Chemistry Vol.88 No.-
Electroporation technique has recently emerged as a tool for delivery of foreign molecules into cells. However, the electroporation has many critical hurdles to overcome in cell viability, delivery efficiency,and productivity. To overcome the hurdles with a single platform, we devised a polyimide (PI)film-basedon- chip electroporation system that shields the cells from the electrodes with four sheathflows,enabling a 3Dflow focusing. This on-chip electroporation with a double forced-flow (OE-DFF)configuration enhances the cell viability to such an extent that even with a long spiral channel for highmolecular delivery efficiency, which is detrimental to the cell viability due to longer exposure to theelectricfield, the cell viability is still increased substantially. The advantages provided by the OE-DFFsystem is demonstrated with afluorescent probe molecule (FITC-BSA) and pPtCrCFP plasmid deliveredinto Chlamydomonas reinhardtii, one of the challenging cell lines to transform. The continuous nature oftheflow system assures high throughput. This novel approach in microfluidic science is expected togreatly contribute to algal research as an efficient electroporation tool as well as to broad applications.
Kim, Sang-Joon,Shin, Hyun-Jae,Kim, Yeu-Chun,Lee, Dae-Sil,Yang, Ji-Won The Korean Society for Biotechnology and Bioengine 2000 Biotechnology and Bioprocess Engineering Vol.5 No.6
Methyl mercaptan oxidase was successfully induced from Rhodococcus rhodochrous IGTS8 using methyl mercaptan gas and purified to homogeneity for the detection of mercaptans. The purification procedure involved DEAE-Sephacel and Superose 12 column chromatography with recovery yields of 85.8 and 83.3%, and a specific activity of 92.7 and 303.4 units/mg-protein, respectively. The molecular weight of purified methyl mercaptan oxidase was determined to be 64.5 kDa by SDS-PAGE. The extract from gel filtration chromatography oxidizes methyl mercaptan to produce formaldehyde, which can be easily detected by the purpald-coloring method. Optimum temperature for activity was achieved at 60$^{\circ}C$. This enzyme was inhibited by both K$_2$SO$_4$and NaCl at concentration of less than 100mM and recovered to original activity at concentration of 200mM. In the presence of methanol, the activity decreased by 33%.
Development of the novel coating formulations for skin vaccination using stainless steel microneedle
Kim, Seong-Jin,Shin, Ju-Hyung,Noh, Jin-Yong,Song, Chang-Seon,Kim, Yeu-Chun Springer-Verlag 2016 Drug delivery and translational research Vol.6 No.5
<P>This study focused on the development of novel coating formulations for stainless steel microneedles against influenza A virus. With in vitro studies, various viscosity enhancers and stabilizers were screened based on the hemagglutination activity of the vaccine, which was coated and dried onto a stainless steel chip at room temperature for 1 day. Following the long-term storage test, the hemagglutination activity and particle size of the vaccine, which was formulated with conventional or methylcellulose or hydroxyethyl cellulose and dried onto the microneedle, were monitored. Next, to evaluate the in vivo immunogenicity and protection effect of each dried vaccine formulation, mice were immunized by the antigen-coated microneedle, which had either the conventional or the proposed formulation. Two novel formulations were chosen in the preliminary screening, and in further evaluations, they exhibited a 20% higher HA activity during storage for 3 months, and no aggregation was observed during storage after drying. In a mouse model, the microneedle with the novel formulation elicited a higher level of IgG and IgG2a was more prevalent in the IgG isotype profile. In addition, mice immunized with the HEC-coated microneedle survived with small weight loss (>90 %) against lethal challenge infection. Overall, the novel formulation hydroxyethyl cellulose preserved significantly higher HA activity during the production and storage of the microneedle as well as improved the in vivo immunogenicity of the vaccine.</P>
Kim, Young Su,Yoon, Nam-kyung,Karisa, Nadia,Seo, Sin-hye,Lee, Jeong-soo,Yoo, Sung-sik,Yoon, In-joong,Kim, Yeu-chun,Lee, Hongweon,Ahn, Jungoh Academic Press 2019 Microbial pathogenesis Vol.127 No.-
<P><B>Abstract</B></P> <P> <I>Streptococcus parauberis</I> is the major infectious agent of streptococcosis in the olive flounder (<I>Paralichthys olivaceus</I>), causing serious economic damage. In this study, we identified potential vaccine candidates against <I>S. parauberis</I> by reverse vaccinology. In total, the 2 out of 21 proteins were identified as vaccine candidates from two available <I>S. parauberis</I> genomes. The membrane-anchored protein SEC10/PgrA and the metal ABC transporter substrate-binding lipoprotein mtsA were potent antigenic proteins based on western blotting with mouse-derived antiserum against whole bacteria of <I>S. parauberis</I> serotypes I and II. In particular, metal ABC transporter substrate-binding lipoprotein (<I>mtsA</I>) showed similar protective immunity to that of whole-cell bacterins against <I>S. parauberis</I> in a zebrafish model. These results suggest that mtsA may be considered as a novel candidate in the development of vaccines against <I>S. parauberis</I>.</P> <P><B>Highlights</B></P> <P> <UL> <LI> We identified 41 vaccine candidates against <I>Streptococcus parauberis.</I> </LI> <LI> SEC10/PgrA and mtsA were potent antigenic proteins. </LI> <LI> <I>mtsA</I> showed high protective immunity in a zebrafish model. </LI> </UL> </P>
Functionalized inclined-GaN based nanoneedles
Kim, Kwon-Ho,Lee, Kyuseung,Hong, Hyeonaug,Yang, Dasom,Ryu, WonHyoung,Nam, Okhyun,Kim, Yeu-Chun THE KOREAN SOCIETY OF INDUSTRIAL AND ENGINEERING 2018 JOURNAL OF INDUSTRIAL AND ENGINEERING CHEMISTRY -S Vol.59 No.-
<P><B>Abstract</B></P> <P>Techniques for the delivery foreign materials into living cells as well as for sensing have undeniable worth in cell biology research. A major barrier for intracellular delivery is to cross the cell membrane with little or no damage. Among the various intracellular delivery methods, one-dimensional nanoneedles are one of the promising methods for insertion with minimal invasiveness to a cell. Most of the previous studies have been conducted using vertical nanoneedle. Here, we synthesized new inclined nanoneedles and demonstrated a novel platform using inclined-gallium nitride nanoneedles (iGaN NNs) to facilitate efficient intracellular delivery. In our system, foreign molecules were successfully delivered into cytoplasm of a cell through the penetration of the cell membrane by the iGaN NNs.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>