肝癌細胞의 γ-Glutamyltranspeptidase에 對한 Monoclonal antibody에 關한 硏究

金明坤,柳總根 고려대학교 의과대학 1992 고려대 의대 잡지 Vol.29 No.2

This study was devised to produce monoclonal antibody against hepatocelullar carcinoma speclfic γ-glutamyltranspeptidase (HCC GGT), which proceeded by first purifying the HCC GGT using ion exchange chromatography and immunoaffinity chromatography. Subsequently, a mouse was immunized with the purified enzyme, and then spleen cells taken from the immunized mouse was fused with mouse myeloma cells (SP2-Ag 14) to form the hybridoma. The following results were obtained by characterization of the monoclonal antibody of the hybridoma using immunoglobulin isotyping and immunoblot. 1. The purification of HCC GGT by means of ion exchange chromatography and immunoaffinity chromatogiaphy produced a yield of 29.41%, a purification fold of 167.87, and final specific activity of 21units per mg of protein. 2. Direct enzyme linked immunoadsorbent assay (ELISA) method was used to measure the antibody level against HCC GGT. An antibody titration of the blood sample taken from the mouse immunized with the above enzyme showed value over 1:5, 120. 3. The cell fusion of the mouse myeloma cell and the spleen cell of the immunized mouse was generated 14 positive wells in 221 wells. (specific fusion efficacy of 6.33%) Only one IgG2a monoclonal antibody was determined by both repetitive cloning procedures and immunoglobulin subisotyplng. 4. A antigen-antibody cross matching after the western blot revealed a specific reaction between the HCC GGT antigen at the 40Kd band and the monoclonal antibody of the hybridoma. And the monoclonal antibody did not react with GGT derived from normal liver or kidney.

단일클론 항γ-Glutamyltransferase항체를 이용한 효소 및 방사 면역측정법 적용에 관한 연구

金明坤,柳總根 고려대학교 의과대학 1994 고려대 의대 잡지 Vol.31 No.1

γ-Glutamyltransferase (GGT: E. C. 2. 3. 2. 2.) is a glycoprotein enzyme which is involved in glutathione metabolism and amino acid transport through the plasma membrane. It is distributed widely in several organs including liver and kidney. Several isozymes of GGT have been reported and some of the isozymes may be associated with hepatocarcinogenesis. We have produced six monoclnal antibodies (mAbs) against GGT purified from the liver of 2-acetamidofluorene (AAF) treated rats. All of the six mAbs were obtained by immunizing mice with liver GGT. Liver GGT from AAF treated rats was biochemically purified to a specific activity of 125.6 units per mg of protein. The overall purification was 551 folds and the final yield was 20. 8. Six hybridomas which produced anti-GGT Abs were extensively subcloned and injected into the peritoneal cavity of BALB/c mice to obtain large quantities of Abs. These mAbs were purified from ascites by ammonium sulfate precipitation and protein A sepharose CL-4B column chromatography. Using these mAbs we performed enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunohistochemistry (IHC), and autoradiography (ARG) to study the distribution of GGT isozymes in tissue. The results indicate that GGT-mAb 1 is specific for the AAF treated liver GGT, GGT-mAb 5 for the normal liver GGT, and GGT-mAb 6 for the normal kidney GGT. These mAbs may be used to evaluate the distribution of GGT isozymes in different tissues.

단일클론 항 γ-Glutamyltrasferase(GCT) 항체를 이용한 혈뇌장벽 내 GGT의 분포에 관한 연구

이병규,김명곤,신규만,류총근 대한신경외과학회 1995 Journal of Korean neurosurgical society Vol.24 No.3

γ-Glutamyltransferase(GGT: E.C. 2.3.2.2.) is a glycoprotein enzyme which is involved in glutathione metabolism and amino acid transport through the plasma membrane. It is distributed widely in several organs including liver, kidney, pancrease and brain. GGTs derived from the brain of Wister rats and BALB/c mice were biochemically purified to a specific activity of 4246.2, 862.1 units per mg of protein, a purification folds 93.7, 43.8 and the final yield 65.8, 44.0% respectively. Electrophoretic pattern of purified GGTs from rats and mice brain shows very similar protein fraction each other. We have produced six monoclonal antibodies(GGT-MAb 1-6) against 2-acetamidofluorene treated rat liver GGT. Using these GGT-MAb 1-6 we performed immunohistochemistry(IHC) to study the distribution of GGY isozymes in normal tissues of rat brain and in ncoplastic tissues of human brain. The results indicated that human brain GGT was localized in pericytes of blood-brain barrier, especially in the blood-rich portion of the brain(e.g. cerebellum of rat, meningioma and craniopharyngioma of human). Therefore these MAbs may be used to evaluate the distribution of GGT isozymes in different tissues.

신경정신 의학분야의 방사성동위원소 표지 cDNA 마이크로어레이

최재걸,신경호,이민수,김명곤 대한핵의학회 2003 핵의학 분자영상 Vol.37 No.1

Microarray technology allows the simultaneous analysis of gene expression patterns of thousands of genes, in a systematic fashion, under a similar set of experimental conditions, thus making the data highly comparable. In some cases arrays are used simply as a primary screen leading to downstream molecular characterization of individual gene candidates. In other cases, the goal of expression profiling is to begin to identify complex regulatory networks underlying developmental processes and diseases states. Microarrays were originally used with cell lines or other simple model systems, More recently, microarrays have been used in the analysis of more complex biological tissues including neural systems and the brain. The application of cDNA arrays in neuropsychiatry has lagged behind other fields for a number of reasons. These include a requirement for a large amount of input probe RNA in fluorescent-glass based array systems and the cellular complexity introduced by multicellular brain and neural tissues. An additional factor that impacts the general use of microarrays in neuropsychiatry is the lack of availability of sequenced clone sets from model systems. While human cDNA clones have been widely available, high quality rat, mouse, and drosophilae, among others are just becoming widely available. A final factor in the application of cDNA microarrays in neuropsychiatry is cost of commercial arrays. As academic microarray facilitates become more commonplace custom made arrays will become more widely available at a lower cost allowing more widespread application. In summary, microarray technology is rapidly having an impact on many areas of biomedical research. Radiosotope-nylon based microarrays offer alternatives that may in some cases be more sensitive, flexible, inexpensive, and universal as compared to other array formats such as fluorescent-glass arrays. in some situations of limited RNA or exotic species, radioactive membrane microarrays may be the most practical experimental approach in studying psychiatric and neurodegenerative disorders, and other complex questions in the brain. (Korean J Nucl Med 2003;37:43-52)

r-Gamma-Glutamyltransferase 의 조직내 분포에 관한 연구 - 단일클론항체의 효소면역측정법 , 방사면역측정법 , 면역조직화학검사 , 자가방사기록검사 적용에 관하여 -

김명곤(Meyoung Kon Kim),박윤규(Youn Kyu Park),류총근(Chong Kun Ryu) 대한핵의학회 1994 핵의학 분자영상 Vol.28 No.1

N/A y-Glutamyltransferase (GGT: E.C. 2.3.2.2.) is a glycoprotein enzyme which is involved in glutath- ione metabolism and amino acid transport through the plasma membrane. It is distributed widely in several organs including liver and kidney. Several isozymes of GGT have been reported and some of the isozymes may be associated with hepatocarcinogenesis. We have produced six monoclnal antibodies (mAbs) against GGT purified from the liver of 2-acetamidofluorene (AAF) treated rats. All of the six mAbs were obtained by immunizing mice with liver GGT. Six hybridomas which produced anti-GGT Abs were extensively subcloned and injected into the peritoneal cavity of BALB/c mice to obtain large quantities of Abs. These mAbs were purified from ascites by ammonium sulfate precipitation and protein A sepharose CL-4B column chromatography. Using these mAbs we proformed enzyme-linked immunosorbent assay (ELISA), radioimmvnoassay (RIA), immunohistochemistry (IHC), and autoradiography (ARG) to study the distribution of GGT isozyme in tissue. The results indicate that GGT-mAb 1 is specific for the AAF treated liver GGT, GGT-mAb 5 for the normal liver GGT, and GGT-mAb 6 for the normal kindey GGT. These mAbs may be used to evaluate the distribution of GGT isozymes in different tissues.

New Liquid Crystal-Embedded PVdF-co-HEP-Based Polymer Electrolytes for Dye-Sensitized Solar Cell Applications

G. Vijayakumar,Meyoung Jin Lee,Myung Kwan Song,Sung Ho Jin,Jae Wook Lee,Chan Woo Lee,Yeong Soon Gal,Hyo Jin Shim,Yong Ku Kang,Gi Won Lee,Kyung Kon Kim,Nam Gyu Park,Suhk Mann Kim 한국고분자학회 2009 Macromolecular Research Vol.17 No.12

New Liquid Crystal-Embedded PVdF-co-HFP-Based Polymer Electrolytes for Dye-Sensitized Solar Cell Applications

Vijayakumar, G.,Lee, Meyoung-Jin,Song, Myung-Kwan,Jin, Sung-Ho,Lee, Jae-Wook,Lee, Chan-Woo,Gal, Yeong-Soon,Shim, Hyo-Jin,Kang, Yong-Ku,Lee, Gi-Won,Kim, Kyung-Kon,Park, Nam-Gyu,Kim, Suhk-Mann The Polymer Society of Korea 2009 Macromolecular Research Vol.17 No.12

Liquid crystal (LC; E7 and/or ML-0249)-embedded, poly(vinylidenefluoride-co-hexafluoropropylene) (PVdF-co-HFP)-based, polymer electrolytes were prepared for use in dye-sensitized solar cells (DSSCs). The electrolytes contained 1-methyl-3-propylimidazolium iodide (PMII), tetrabutylammonium iodide (TBAI), and iodine ($I_2$), which participate in the $I_3^-/I^-$ redox couple. The incorporation of photochemically stable PVdF-co-HFP in the DSSCs created a stable polymer electrolyte that resisted leakage and volatilization. DSSCs, with liquid crystal(LC)-embedded PVdF-co-HFP-based polymer electrolytes between the amphiphilic ruthenium dye N719 absorbed to the nanocrystalline $TiO_2$ photoanode and the Pt counter electrode, were fabricated. These DSSCs displayed enhanced redox couple reduction and reduced charge recombination in comparison to that fabricated from the conventional PVdF-co-HFP-based polymer electrolyte. The behavior of the polymer electrolyte was improved by the addition of optimized amounts of plasticizers, such as ethylene carbonate (EC) and propylene carbonate (PC). The significantly increased short-circuit current density ($J_{sc}$, $14.60\;mA/cm^2$) and open-circuit voltage ($V_{oc}$, 0.68 V) of these DSSCs led to a high power conversion efficiency (PCE) of 6.42% and a fill factor of 0.65 under a standard light intensity of $100\;mW/cm^2$ irradiation of AM 1.5 sunlight. A DSSC fabricated by using E7-embedded PVdF-co-HFP-based polymer electrolyte exhibited a maximum incident photon-to-current conversion efficiency (IPCE) of 50%.

Pharmacokinetics of 111In-labeled Triplex-forming Oligonucleotide Targeting Human N-myc Gene

Jae Gol Choe,Meyoung-Kon Kim,Eun Jung Oh,Eun Jeong Yoon,손정원,Min Kyu Park,Gil Hong Park 한국분자세포생물학회 2002 Molecules and cells Vol.14 No.1

Environmental Pollution and Gene Expression: Dioxin

Kim, Ki-Nam,Kim, Meyoung-Kon The Korean Society of Toxicogenomics and Toxicopro 2005 Molecular & cellular toxicology Vol.1 No.2

Dioxins, especially 2, 3, 7, 8-Tetrachlorodibenzo-p-dioxin (TCDD or dioxin), are ubiquitous environmental contaminants. TCDD is known that it has toxic effects in animals and humans, including chloracne, immune, reproductive and developmental toxicities, carcinogenicity, wasting syndrome and death. TCDD induces a broad spectrum of biological responses, including disruption of normal hormone signaling pathways, reproductive and developmental defects, immunotoxicity, liver damage, wasting syndrome and cancer. Many researches showed that TCDD induces gene expression of transcriptional factors related cell proliferation, signal transduction, immune system and cell cycle arrest at molecular and cellular levels. These toxic actions of TCDD are usually mediated with AhR (receptor, resulted from cell culture, animal and clinical studies). cDNA microarray can be used as a highly sensitive and informative marker for toxicity. Additionally, microarray analysis of dioxin-toxicity is able to provide an opportunity for the development of candidate bridging biomarkers of dioxin-toxicity. Through microarray technology, it is possible to understand the therapeutic effects of agonists within the context of toxic effects, classify new chemicals as to their complete effects on biological systems, and identify environmental factors that may influence safety.

Promising Next Generation Technology in Toxicology-Toxicogenomics

Ryu, Jae-Chun,Kim, Meyoung-Kon,Cho, Man-Ho,Chun, Tae-Hoon The Korean Society of Toxicogenomics and Toxicopro 2005 Molecular & cellular toxicology Vol.1 No.1

Toxicology is a multidisciplinary field, and an important science that impacts both environmental health regulation and the development and practice of medicine. The rapid progress in cellular and molecular biology, like many other branches of biomedical research, toxicology is now experiencing a renaissance fueled by the application of "omic" technologies to gain a better understanding of the biological basis of toxicology of drugs and other environmental factors. In this review on current progress on toxicology, the future perspective, concept, approaches and applications of toxicogenomics as next generation promising technology in toxicology field will be described.