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.

서울 시내 4년제 남자 대학생의 우울과 흡연량, 흡연기간, 니코틴 의존도와의 관계

박지영,박소영,이미숙,백수진,신의경,예혜련,오승진,장윤정,조혜진,김수지 이화여자대학교 간호과학대학 2003 이화간호학회지 Vol.- No.37

Background: With reports on the relation between smoking and depression appearing since 1980, it has become clear that depression is one of the psychodynamics of smoking, though there has been little or no progress made in the study of whether or not there is indeed a relation between depression and nicotine dependence. Purpose: The purpose of the study was to investigate the effects of depression on the amount and period of smoking and nicotine dependence among university man student smokers. Method: In periodical examination between 14 and 21, May, 2003, using 170 university male. The CES-D(Center for Epidemiologic Studies Depression) Scale and the FTQ(Fagerstrom Tolerance Qustionnaire) were utilized to measure the level of depression and nicotine dependency. The amount of smoking was calculated based on the number of cigarette used. Result: Results indicated that depression had something to do with amount of smoking. Nicotine dependence also was related to depression but total value was relatively low. finally the period of smoking had nothing to do with depression. Conclusion: The results of this study revealed that the amount of smoking of university man student smokers was very high. Both the amount of smoking and nicotine dependence were affected by levels of depression. Accordingly psychiatric nursing access such as emotional support is needed to manage depression and decrease smoking.

HSS STI-CMP적용을 위한이중 패드의 최적화

서용진,박성우,김철복,정소용,이경진,김기욱,박창준 대불대학교 2002 大佛大學校大學院 硏究論文集 Vol.- No.1

As the device geometry shrinks to the deep submicron region, chemical mechanical polishing(CMP) planarization become a more essential technique of advanced ULSI process. Also, CMP process was required for the global planarization of inter-metal dielectric(IMD), inter-level dielectric(ILD) layers and interconnections with free-defect. Especially, the complete global planarization of IMD, ILD and interconnections can be achieved only with the CMP process. However, as the IMD and ILD layer gets thinner, several problems were found in the CMP process. It does have various problems such as dishing effect, torn oxide defects and nitride residues in oxide. So, it leads to severe circuit failure, which affects yield. In this paper, we studied the characteristics of polishing pad, which can apply STI-CMP process for global planarization of multilevel interconnection structure. Also, we investigated the effects of different sets of polishing pad, such as soft and hard pad. As an experimental result, hard pad showed center-fast type, and soft pad showed edge-fast type. Totally, the defect level has shown little difference, however, the counts of scratch was defected less than 2 on JR111 pad. Through the above result, we can select optimum polishing pad, so we can expect the improvement of throughput and device yield.

林地利用에 따른 溪流水의 水質變化

具瀟瑛,鄭源玉,金弘洙,朴晋源,麻鎬燮 경상대학교 농과대학 부속연습림 2001 演習林硏究報告 Vol.- No.11

Table 2. Evaluation of stream water quality in three stands by Drinking water quality standard <원문참조> 임지이용에 따른 수질변화 및 오염정도를 구명하여 향후 비점오염원 관리를 위한 기초자료를 제공하고자 경남 진주시 정평리에 위치한 3개 임분(소나무, 밤나무Ⅰ, 반나무Ⅱ임분)에서 2000년 7월부터 9월까지 계류수의 수질변화를 분석하여 얻은 결과는 다음과 같다. 1. 세 임분의 계류수 pH는 소나무 임분(pH6.59)>밤나무 Ⅱ임분(ph 6.53)>밤나무 Ⅰ임분(pH 6.47)계류수 순으로 나타났다. 2. 전기전도도는 밤나무 Ⅰ임분>소나무 임분>밤나무 Ⅱ임분 계류수의 순으로 높게 나타났다. 3. 양이온은 세 임분의 계류수 모두 Ca^2+, Na^+, Mg^2+, K^+, NH_^+의 순으로 높게 나타났다. 4. 음이온은 소나무 임분과 밤나무 Ⅱ임분 계류수에서는 SO_4^2, Cl, NO_3의 순으로 높게 나타났고, 밤나무 Ⅰ임분 계류수에서는 NO_3, SO_4^2, Cl 의 순으로 높게 나타나 차이를 보였다. 5. 소나무, 밤나무 Ⅱ임분 계류수는 pH, NH_4+, NO_3, Cl, SO_4^2이 먹는 물 수질기준 범위내에 있었다. 그러나 밤나무 Ⅰ임분 계류수에서 5항목 중 NO_3는 먹는 물 수질기준을 넘어서는 범위에 있어 임지이용에 따른 하류수질의 오염이 우려되었다. 따라서 이러한 상류지역에서 오염원 차단을 위한 수질정화시설 등의 대책이 마련되어야 할 것으로 사료된다. This study was carried out to clarify the change characteristics of stream water quality by land-use of forest from July to September, 2000 in three stands(Pinus densiflora, Castanea crenata(Ⅰ), Castanea crenata(Ⅱ) stand) of Jeongpyeong-ri, Jinju-si, Gyeongnam. The mean pH of rainfall results in acid rain of 5.3. The pH of stream water in three stands was hugh in order of Pinus densiflora (pH 6.59), Castanea crenata(Ⅱ)(pH 6.53) and Castanea crenata(Ⅰ) stand(pH 6.47). The electrical conductivity of stream water was high in order of Castanea crenata(Ⅰ), Pinus densiflora and Castanea crenata(Ⅱ) stand. Cations of three stand in stream water high in order of Ca^2+, Na^+, Mg^2+, K^+ and NH_4^+. But anions of stream water in Pinus densiflora stand and Castanea crenata(Ⅱ) stand were high in order of SO_4^2, Cl and NO_3. In Castanea crenata(Ⅰ) stand cations of stream water were high in order of NO_3, SO_^2 and Cl. The level of pH, NH_4^+, NO_3, Cl and SO_4^2 of stream water in Pinus densiflora stand and Castanea crenata(Ⅱ) stand reached within the level of domestic use standard for drinking water But the level of NO_3 of stream water in Castanea crenata(Ⅰ) stand was higher than that of domestic use standard. Therefore, non-point sources like forest watersheds which are fertilizer application lands should be taken to the appropriate mitigation measures.

캐런 헤세의 『모래 폭풍을 지나서』를 통해 본 생명의 공존성 회복의 비전

박소진(Park So-Jin) 문학과 환경학회 2010 문학과 환경 Vol.9 No.1

This paper examines the close connections between nature and human beings, between human beings in a community, and between human beings’ physical condition and their emotional state, through Karen Hesse’s book, Out of Dust (1997). It describes the harsh reality that farmers in Oklahoma, which was caused by the combination of the economic depression and the most destructive ‘natural disaster’ in the history of the USA, the so-called Dust Bowl. The Dust Bowl resulted from human abuse of the farm plains, the removal of the grass, and the destruction of the natural environmental balance in the plains. This novel, written in the form of a diary, portrays how destructive the dust and drought were during that period, and how devastated farmers’ lives were, focusing on one main character, Billy, her parents, and their neighbors. While this book warns against people’s greed for short term profit and shows their blindness concerning the consequent disaster, it also demonstrates what makes people overcome those hardships, and how the main characters sustain their hope for life despite bitter mistakes and conflicts. Hope is found in the comfort derived from the forces of nature and from people's love and forgiveness for each other, even though nature and people are the very elements that cause the misery and scars in their lives. Nature and people are portrayed in contrasting ways. No matter how destructive and threatening nature is, it is a fundamental influence on people’s lives, and no matter how unforgivable and disappointing people (especially family members) are, they may be a source of great support. Suggesting the importance of the interdependence between nature and human beings, and also between human beings, this book describes these connections by drawing parallels between these four aspects: 1) the state of nature, 2) the state of people’s lives, 3) people’s physical and emotional condition, and 4) people’s relationships. This book implies that nature, human beings’ physical and emotional condition, and the health of the community, are closely related to each other. This is related to an ecological perspective, which perceives the relationship between nature and human beings as an organic whole, no longer regarding them as separate entities.

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.

Interacelluar reprogramming of expression, glycosylation, and function of plant-derived anti-virus therapeutic monoclonal antibody

Da-Young Park,Kyung-Jin Lee,Yangkang So,Zhe Lu,Seulki Lee,Jung-Hwan Lee,Seung Han Oh,Yeon Soo Han,Doo-Byoung Oh,Youngkwan Kim,Kisung Ko 한국당과학회 2010 한국당과학회 학술대회 Vol.2010 No.1

Plant genetic engineering has led to production of plant-derived mAb (mAbP), which can provides a safe and economically effective alternative to the conventional antibody expression methods. In this study, the expression levels of mAbP SO57 with or without ER-retention peptide extensions signal (Lys-Asp-Glu-Leu; KDEL) in transgenic tobacco plants were analysed in transgenic plant. The expression levels of mAbP SO57 with KDEL were significantly higher than that without KDEL regardless of transcription level. mAbP SO57 with KDEL localized surround to the nucleus suggesting that the mAbP with KDEL is localized in ER. The mAbP without KDEL and mAbH had mainly Golgi type glycans, whereas the ER-localized mAbP with KDEL had glycan profile with both oligomannose type (47.6%) and Golgi type (52.4%). The Fc domains of both purified mAbP (with and without KDEL) and human-derived mAb (mAbH) had similar bingding activity to the Fcγ RI receptor (CD64). Both mAbP (with and without KDEL) had a shorter half-life than mAbH. However, both mAbP with and without KDEL was as effective at neutralizing activity of the rabies virus CVS-11 as the mAbH. These results suggest that ER localization of recombinant mAbP by KDEL reprograms glycosylation and enhances production of the functional antivirus therapeutic antibody in the plant.

신경교세포 및 RAW 264.7 세포에서 Protein kinase의 활성에 의한 유도성 Nitric oxide synthase의 발현

박상철,노삼길,배소현,박지선,이충재,허강민,석정호,이재흔 충남대학교 의학연구소 2003 충남의대잡지 Vol.30 No.2

NO(nitric oxide) plays an important role as neurotransmitter or cytokine, and pathologic factor for some diseases by the large amount production with iNOS(inducible NO synthase) expression in macrophages or glial cells. The expression of iNOS is regulated by various cytokines, protein kinases and transcription factors. In this experiment, to investigate the roles of progein kinase and NF-kB for iNOS expression, the effects of PMA(phorbol 12-myristate 13-acetate), cAMP, and various protein kinase inhibitors on LPS(lipopolysaccharide)-induced iNOS mRNAN expression and nuclear NF-kB binding complex were examined in C6 glial cells and RAW 264.7 cells. In C6 glial cells, iNOS mRNA expression by LPS was induced from 1 hour and peak at 3 hour after treatment. In RAW 264.7 cells, the mRNA was observed from 3 hour and peak at 6 hour. PMA enhanced markedly LPS-induced iNOS mRNA expression and NF-kB binding complex in C6 glial cells, but did not much influence on LPS-induced iNOS mRNA expression in RAW 264.7 cells, in spite of increased LPS-induced NF-kB binding complex at 30 min. cAMP(dibutyryl cAMP) did not much influence on LPS-induced iNOS mRNA expression, by increased LPS-induced NF-kB binding complex in C6 glial cells. However, in RAW 264.7 cells, cAMP increased slightly LPS-induced iNOS mRNA expression without change of NF-kB binding complex. Staurosporine did not influence on LPS-induced iNOS mRNA expression and NF-kB binding complex in C6 glial cells, but in RAW 264.7 cells, decreased LPS-induced iNOS mRNA expression and NF-kB binding complex. Ro-31-8220 did not much influence on LPS-induced iNOS mRNA expression and NF-kB binding complex in C6 glial cells, but in RAW 264.7 cells, decreased significantly LPS-induced iNOS mRNA expression in spite of increased LPS-induced NF-kB binding complex for 3hours. G 6976 did not much influence on LPS-induced iNOS mRNA expression with decreased NF-kB binding complex in C6 glial cells, but in RAW 264.7 cells, decreased iNOS mRNA expression without influence on LPS-induced NF-kB binding complex. Genistein did not influence on LPS-induced iNOS mRNA expression and NF-kB binding complex in C6 glial cells, but in RAW 264.7 cells, decreased LPS-induced iNOS mRNA expression inspite of increased NF-kB binding complex. These results suggest that LPS-induced regulation of iNOS expression or NF-kB activity in C6 glial cells, might be different from RAW 264.7 cells through various protein kinases or other factors.

Headspace Sampler와 Gas Chromatography를 이용한 방향족 화합물의 무차원 헨리 상수 측정

박수진,임화윤,박소진 한국화학공학회 2004 Korean Chemical Engineering Research(HWAHAK KONGHA Vol.42 No.1

방향족화합물의 무차원 헨리상수(Air/Water 분배계수 : K_(aw))를 headspace sampler(HSS)와 gas chromatography(GC)를 이용하여 318.15-355.15 K 온도 범위에서 온도를 변화시키며 측정하였으며, 이 방법으로 시료와 평형 및 채취, 분석 과정에 있어서 용질의 휘발로 인한 오차와 수작업에 의한 측정 오차를 최소한으로 줄이고자 하였다. HSS내 평형용기의 기상과 액상조성은 GC의 피크면적과 물질수지 식으로부터 계산하였으며, 이 관계식을 이용하여 benzene을 비롯한 몇 개의 방향족 화합물의 무차원 헨리상수를 온도별로 측정하였고 문헌값과 비교하여 방법의 신뢰성을 확인하였다. 또한 온도와 무차원 헨리상수의 함수관계를 통해 온도에 따른 무차원 헨리상수 값을 유추하는 방법의 가능성을 살펴보았다. Dimensionless Henry's law constants of aromatic compounds in water were measured at various temperatures between 318.15-355.15K using a headspace autosampler with a gas chromatography. This measuring system prevent, sample losses due to volatilization from equilibration, sampling, and chemical concentration analysis by a high precision headspace sampler and gives relatively accurate results. Dimensionless Henry's law constant is calculated by thermodynamic relation and the mass balance with gas chromatographic peak area. Different equilibrium concentration in gas phase was made by using different gas-liquid volume ratio in sample vial. The temperature dependence of dimensionless Henry's law constant was correlated with the classical van't Hoff equation as a function of temperature. The extrapolated data at 298.15K from correlated equation were compared with available literature data in order to test this method.

김미료 소재로서 Di-D-Fructofuranose Dianhydride3의 물리 및 생리적 특성

박정복,김소자,최용진 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.5

충치 원인균인 Streptococcus mutans와 유용 장내 세균인 각종 Bifidus 균주 증식에 미치는 di-D-fructofuranose dianhydrideIII(DFAIII)의 효과를 조사 분석하여 DFAIII의 생리 효과를 간접적으로 분석하였다. DFAIII는 Arthrobacter sp. A-6 토양 분리균이 생산하는 inulin fructotransferase(depolymerizing)을 이용, inulin을 분해하여 다량 생산한 다음 gel filtration 과정을 거쳐 순수 분리 정제하였다. 정제 DFAIII는 pH 2.5, 130℃에서 15분 가열 처리에 의해 약 35%의 분해율을 보이는 반면 sucrose는 완전히 분해되었으며, pH 5.5, 140℃ 처리 조건에서는 거의 분해되지 않는 매우 높은 열 안정성을 나타내었다. 또한 충치 유발의 중요 기준이 될 수 있는 S. mutans 증식 효과도 fructose와 sucrose는 물론이고 시판 올리고당보다도 훨씬 낮았다. 반면, Bifidus 균에 대한 증식 효과는 균주에 따라 다소의 차이는 있었으나 증식을 전혀 보이지 않았던 E. coli에 비해 뚜렷한 증식 촉진효과를 나타내었다. BL 배지에서의 Bifidus longum과 E. coli의 혼합 배양에 있어서도 Bifidus longum의 경우에서는 선택적으로 뚜렷한 증식 촉진 효과를 보였으나 E. coli의 증식은 오히려 억제됨을 확인하였다. 이상의 실험 결과를 종합 분석해 본 결과 DFAIII는 매우 우수한 생리 효과를 기대할 수 있는 감미료 소재중의 하나인 것으로 평가되었다. Some physical and physiological properties of di-D-fructofuranose dianhydrideIII(DFAIII), as a new sweetener, were investigated via in vitro experiments. The disaccharide was prepared by decomposing inulin with inulin fructotransferase(depolymerizing) from Arthrobacter sp. A-6. DFAIII had more excellent heat and acid stability than sucrose. This was one of the most desirable properties especially for the oligomer types of sweetener. DFAIII showed the least pH drop in the Streptococcus mutans culture, compared with the other saccharides examined. This indicates that the sugar will be fairly effective for prevenreas, E. coli did not show growth promotion in the DFAIII-containing medium. In the co-culture of Bifidus longum and E. coli in the BL medium, Bifidus longum had a selective growth while the growth of E. coli appeared rather to be inhibited.