Effect of ER signal peptides (KDEL) on expression and function of anti-virus monoclonal antibody in plant

Da-Young Park,Yangkang So,Kyung-Jin Lee,Zhe Lu,Eun-Yi Oh,Kwang-Wok Min,Seul-Ki Lee,Jung-Hwan Lee,Mi-Hyun Ahn,Mira Song,Doo-Byoung Oh,Youngkwan Kim,Kisung Ko 한국당과학회 2009 한국당과학회 학술대회 Vol.2009 No.1

Plant genetic engineering has led to the production of plant-derived mAb (mAbP), which provides a safe and economically feasible alternative to the current antibody expression systems. In this study, the expression levels of mAbP SO57 with or without ER-signal peptides (Lys-Asp-Gly-Leu;KDEL) in transgenic tobacco plants were analysed in transgenic plant. PCR and Reverse Transcription-PCR analyses showed existence of heavy and light chain genes of mAb with or without KEDL and their transcription in plant, respectively. Western blot showed that the expression levels of mAbP SO57 with KDEL were significantly higher than that without KDEL. Flow cytometry analysis showed that the Fc domains of both purified mAbP and mammalian-derived mAb have similar binding activity to the FcγRI receptor. High-performance liquid chromatography analysis showed that the mAbP SO57 with KDEL had glycan profile with both oligomannose and golgi type, whereas the mAbP SO57 without KDEL had only golgi type glycans. Neutralizing analysis with rabies virus CVS-11 showed the similar neutralizing activity between mAbP SO57 with and without KDEL. These results suggest that the potential of mAbP SO57 for rabies immunotherapy is regardless of plant specific glycan structures.

Effect of ER signal peptides (KDEL) on expression and function of monoclonal antibody in plant

Da-Young Park,Eun-Yi Oh,Zhe Lu,Yangkang So,Mi-Hyun Ahn,Mira Song,Kyung-Jin Lee,Doo-Byoung Oh,Youngkwan Kim,Kisung Ko 한국당과학회 2009 한국당과학회 학술대회 Vol.2009 No.1

Plant genetic engineering led to the production of plant-derived mAb (mAbP), which provides a safe and economically feasible alternative to the current animal expression systems. In this study, the expression level of mAbP SO57 between with and without Lys-Asp-Glu-Leu (KDEL) was compared in transgenic plant. PCR and PT-PCR analyse showed stable gene insertion transcription of heavy and light chain genes of mAbP SO57 with or without KDEL in plant, respectively. We validated expression of mAbP SO57 by western blot. Western blot showed the significantly higher expression level of mAbP SO57 with KDEL compared to without KDEL. Flow cytometry (FACS) analysis showed that the Fc domains of both purified mAbP and mammalian-derived mAb (mAbM) evidenced similar binding activity to the FcγRI receptor (CD64). High performance liquid chromatography (HPLC) analysed, glycosylation patterns of mAbP SO57 with or without KDEL. The mAbP SO57 with KDEL had glycan profile with both oligomannose type (47.6%) and golgi type (52.4%), while the mAbP SO57 without KDEL had only golgi type (100%) glycans. Neutralizing analysis with rabies virus CVS-11 showed the similar neutralizing activity between mAbP SO57 with and without KDEL. These results suggest the potential of mAbP SO57 for rabies immunotherapy, regardless of plant specific glycan structures.

교내 실내 공기에서 부유 미생물의 분포

박창은,임미소,이다은,김설아,강재원 남서울대학교 2010 남서울대학교 논문집 Vol.16 No.2

The pathogenic and non-pathogenic microorganism have been isolated from the indoor airborne, the closest environment of human beings. Our purpose were to investigate the distribution of fungi and bacteria in indoor airborne sampled from 7 locations in Namseoul University for 2 months from September to October, 2010. The isolated bacteria were identified by Gram stain and biochemical test using VITEKTM system, also The fungus were identified by LPCB. The experiment on microorganism concentration of indoor air carried out and the average of total microorganism was measured. When airborne bacteria were isolated and identified. The isolation rates of Gram positive cocci, Gram positive bacilli, Gram negative bacilli, and Gram negative cocci were 38.3%, 22.4%, 2.7%, 0%, respectively. In Gram positive cocci, the most strains were identified as Micrococcus spp. In Gram positive bacilli, the most strains were identified as Bacillus spp. The frequently isolated fungi were Aspergillus sp, Penicillium sp, Alternaria sp, Rhizopus sp, Mucor sp. respectively. Further systematic studies are necessary with an emphasis on species identification. Key Words : Microorganism, Fungus, Indoor Airborne, Micrococcus, Aspergillus

교내 수도꼭지형 냉온수기의 생수 배출 부위와 생수에 존재하는 미생물의 분포

박창은,김설아,임미소,이다은,박유진 남서울대학교 2010 남서울대학교 논문집 Vol.16 No.1-1

The distributed species and number of viable microorganisms in drinking water were monitored according to water dispenser maintenance. our purpose was to investigate the distribution of bacteria in drinking water dispenser. To analyze possible dispenser contaminations sampling were performed from 14 locations in Namseoul University for 3 months from March to May, 2010. The isolated bacteria were identified by Gram stain and biochemical test using VITEKⅡ systems. The microorganism concentrations around the water discharge areas were measured and the average of total microorganisms was determined. The number of bacteria were measured by McFarland turbidity. The water and water discharge areas were separated and identified. The isolation rates of Gram positive cocci and Gram negative bacilli were 56.5%, 17.4% respectively. The frequently isolated bacteria were Staphylococcus epidermidis, Enterobacter aerogenes, Enterbacter sakazakii, Alcaligenes faecalis, Acinetobacter baumannii, Staphylococcus hominis, respectively. Further systematic studies are necessary with an emphasis on species identification. Key Words : Microorganisms, Water Discharge Areas, Drinking Water, McFarland

Perspectives on the systematic review for the 2020 Dietary Reference Intakes for Koreans for calcium

So Young Bu,Mi Ja Choi,Da Seul Choi,You-Mi Jung,In-Sil Jang,Narae Yang,Kirang Kim,Clara Yongjoo Park 한국영양학회 2022 Nutrition Research and Practice Vol.16 No.no.sup1

An accurate assessment of the recommended calcium (Ca) intake may contribute to reducing the risk of fractures and chronic diseases, ultimately improving quality of life. This review was performed to summarize key findings of Ca studies, investigate the effect of Ca intake on health outcomes, and determine the adequacy of evidence to revise the 2015 Dietary Reference Intakes for Koreans (KDRIs) for Ca in 2020. Databases were searched for intervention studies that assessed health outcomes by providing Ca in diets or as supplements. The framework of the systematic review comprised conducting literature searches, data extraction, quality assessment of the literature, and summarizing key findings relevant to set the Estimated Average Requirement (EAR) and Tolerable Upper Intake Level (UL) for Ca for the 2020 KDRI. The final search was performed in June 2019. A total of 13,309 studies were identified through databases and manual search. Sixtyfive studies were included in the final quality assessment and were summarized according to health indicators. As bone health was used as an indicator of the EAR for Ca, literature reports on bone health were further categorized by the life-cycle stage of the participants. This systematic review did not find new evidence that could be applied to the general Korean adult population, including postmenopausal women, for defining a new EAR for Ca in the 2020 KDRIs. Evidence in most of the reviewed literature was considered weak; however, some evidence was found that could improve the criteria on how the EAR for Ca was determined in children and adolescents. A review of the literature for the 2020 KDRIs for Ca did not find strong evidence in order to change the recommended values of the 2015 KDRIs. More clinical interventions are required among Koreans to strengthen the body of evidence to warrant the revision of the KDRIs.

Crizotinib attenuates cancer metastasis by inhibiting TGFβ signaling in non-small cell lung cancer cells

Park Soonbum,Cho Eun A,Chun Jung Nyeo,Lee Da Young,Lee Sanghoon,Kim Mi Yeon,Bae Sang Mun,Jo Su In,Lee So Hee,Park Hyun Ho,김태민,So Insuk,Kim Sang-Yeob,Jeon Ju-Hong 생화학분자생물학회 2022 Experimental and molecular medicine Vol.54 No.-

Crizotinib is a clinically approved tyrosine kinase inhibitor for the treatment of patients with locally advanced or metastatic non-small cell lung cancer (NSCLC) harboring EML4-ALK fusion. Crizotinib was originally developed as an inhibitor of MET (HGF receptor), which is involved in the metastatic cascade. However, little is known about whether crizotinib inhibits tumor metastasis in NSCLC cells. In this study, we found that crizotinib suppressed TGFβ signaling by blocking Smad phosphorylation in an ALK/MET/RON/ROS1-independent manner in NSCLC cells. Molecular docking and in vitro enzyme activity assays showed that crizotinib directly inhibited the kinase activity of TGFβ receptor I through a competitive inhibition mode. Cell tracking, scratch wound, and transwell migration assays showed that crizotinib simultaneously inhibited TGFβ- and HGF-mediated NSCLC cell migration and invasion. In addition, in vivo bioluminescence imaging analysis showed that crizotinib suppressed the metastatic capacity of NSCLC cells. Our results demonstrate that crizotinib attenuates cancer metastasis by inhibiting TGFβ signaling in NSCLC cells. Therefore, our findings will help to advance our understanding of the anticancer action of crizotinib and provide insight into future clinical investigations.

Expression of Anti-colorectal Cancer Monoclonal Antibody CO17-1A in Insect Cells.

Mi-Hyun Ahn,Mira Song,Kyung-Jin Lee,Eun-Yi Oh,Zhe Lu,Da-Young Park,Yang-Kang So,Doo-Byoung Oh,Kisung Ko 한국당과학회 2008 한국당과학회 학술대회 Vol.2008 No.1

Insect cell expression system using baculovirus has several benefits from high capacity, flexibility, safety to humans and glycosylation capability. Thus, the baculovirus insect cell system has been widely used for production of recombinant protein. In this study, we designed the insect cell expression system to produce anti-cancer mAb CO17-1A, which recognizes the antigen GA733 highly expressed on the surface membrane of colorectal carcinoma cells. The heavy (HC) and light chain (LC) genes of the mAb CO17-1A were cloned under the control of two different promoters, P10 and PPH, respectively on baculovirus expression, pFastBacTM Dual vector. The gene expression cassettes carrying HC and LC genes were transfered into a parent bacmid in E.coli (DH10Bac). The bacmid was transfected to Sf9 insect cells to generate the baculovirus expressing mAb CO17-1A (CO17-1A-Bac virus). Western blot and immunoflouorescence confocal analyses confirmed the mAb CO17-1A expression in CO17-1A-Bac virus infected insect cells. The optimum conditions for mAb expression were validated at 24, 48 and 72h after the virus infection with MOI (optimum virus concentration) ranging (0.2, 1 and 5). Expression of mAb CO17-1A in insect cells significantly increased at both 48 and 72h after transfection with the MOI 1. HPLC chromatography revealed that the mAb CO17-1A expressed in the insect cell had insect specific glycan structures. Cell ELISA showed that the purified mAb from insect cell cultured media had a specific binding activity to SW948 human colorectal cancer cell. These results indicated that the baculovirus insect cell system is able to express, assemble, and secrete functional full size monoclonal antibody with insect specific glycosylation.

Expression of Anti-Colorectal Cancer Immunotherapeutic Monoclonal Antibody in Transgenic Plant

Yangkang So,Zhe Lu,Eun-Yi Oh,Da-Young Park,Kyung-Jin Lee,Mira Song,Seung Han Oh,Arshad Jamal,Mi-Hyun Ahn,Yeon Soo Han,Doo-Byoung Oh,Youngkwan Kim,Kisung Ko 한국당과학회 2009 한국당과학회 학술대회 Vol.2009 No.1

The mAb CO17-1A recognizes the colorectal antigen GA733-2E, which highly is expressed on the surface membrane of human colorectal carcinoma cells. We have successfully developed plant system for production of anti-colorectal cancer monoclonal antibody, mAb CO17-1A. Plant expression system for mAb production has several advantages in terms of the lack of animal pathogenic contaminants, low cost, ease of scale-up for agricultural production, and efficient post-translational glycosylation modifications compared to the conventional fermentation methods. KDEL (Lys-Asp-Glu-Leu), ER retention signal attached to C-terminus of protein might induce high protein expression and the glycoprotein carrying oligomannose glycans. In this study, the expression level of mAb CO17-1A was compared between mAbs fused with and without KDEL in plant. PCR analysis showed stable gene insertion of heavy and light chain genes of mAb CO17-1A with or without KDEL. We validated the expression of mAb CO17-1A by western blot and its subcellular localization under the confocal microscopy. Western blot showed the significantly increased expression level of mAb CO17-1A with KDEL compared to without KDEL. Cell ELISA with human colorectal carcinoma cell line confirmed that expression of the mAb with KDEL was significantly higher than that of mAb without KDEL. The glycosylation analysis revealed that mAb CO17-1A without KDEL had plant specific glycans structure while mAb CO17-1A with KDEL had mainly oligomannose type glycans. Taken together, these studies suggest that KDEL affects expression of mAb and its glycan structures.

자체 제작 유방 진단 팬텀과 타사 팬텀의 비교

최다영(Da-Young Choi),이단비(Dan-Bi Lee),이소연(So-Yeon Lee),이지윤(Ji-Yoon Lee),전수빈(Su-Bin Jeon),정미애(Mi-Ae Jung),박지혜(Ji-Hye Park),김재현(Jae?Hyeon Kim) 대한초음파의료영상학회 2018 대한초음파의료영상학회 학술대회 초록집 Vol.3 No.1

근린의 아동 놀이환경과 놀이행태 연구설계의 특성

박소현(Park, So-Hyun),유다은(Yoo, Da-Un),최이명(Choi, Yee-Myung),강현미(Kang, Hyun-Mi),김현주(Kim, Hyun-Ju) 대한건축학회 2015 대한건축학회논문집 Vol.31 No.12

The purpose of this study is to review recent causality studies on relationship between children’s play environment and play behavior in residential neighborhoods. Seven key precedent studies in the past ten years have been analyzed to understand their research design structures and procedures: research objectives, research subjects & scope, and research methods. The research findings are as follows. First, complement of research design is required that both qualitative and quantitative methods are connected for examining definitive relationship between children’s play environment and play behavior. Second, showing characteristics of children"s play behavior, researches for children’s play environment are differentiated itself from existing walkability studies of residential neighborhoods. In addition, This study highlights critical issues and methodological commonalities shared among researchers in disciplines of public health, transportation, urban planing, etc. The results of this study provides preliminary foundations and guidelines for future researches of the physical environment concerning children"s outdoor play.