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보건의료 R&D 연구성과 활용·확산을 위한 특허맵 분석 및 활용방안 연구
전수환(Su-Hwan Cheon),이경민(Kyung-Min Lee),김미경(Mi-Kyoung Kim),제영태(Young-Tae Je),신상훈(Sang-Hun Shin),김명환(Myung-Hwan Kim),김동석(Dong-Seok Kim),박성호(Seong-Ho Park),김기택(Gi-Tae Kim),곽정애(Jung-ae Kwak),전혜경(Hye-Kyoung 한국생물공학회 2013 KSBB Journal Vol.28 No.6
Translational research (TR) as high quality research can accelerate collaboration strongly between biotechnology-based researchers and clinical-research experts for overcoming diseases. TR facilitates basic science translated to clinical efficacy and effectiveness from bench (basic science) to bedside (clinical practice) for the enhancement of human health. Disease-oriented TR programs were defined as unilateral, bilateral and multilateral TR in this patent performance analysis. Patent performance was measured in a R&D project on Health and Medical Technology to enhance the productivity of R&D investment on disease-oriented TR in Health Technology (HT). Patent Map (PM) analysis and Bibliometrics were conducted to collect information for the assessment of research patents of TR programs. Futhermore, PIAS (Patent Information Analysis System) and Thinklear programs were applied for quantitative and qualitative analysis successfully. These indicate that multi-dimensional analysis of patent performance for disease-oriented TR could promote the connection of R&D-IP (Research and Development-Intellectural Property) and R&BD (Research and Business Development) supporting system significantly.
생명공학기술적 관점에서 질병중심 중개연구의 효율적 성과분석에 대한 실증연구
전수환(Su-Hwan Cheon),정성철(Sung-Chul Jung),제영태(Young-Tae Je),김기태(Gi-Tae Kim),김명환(Myung-Hwan Kim),박성호(Seong-Ho Park),전혜경(Hye-Kyoung Jeon),권준영(Jun-Young Kwon),김동일(Dong-Il Kim),김동석(Dong-Seok Kim),이경민(Kyung-M 한국생물공학회 2012 KSBB Journal Vol.27 No.1
Recently, translational research (TR) in health technology (HT) has been considered as an emerging alternative research system for the improvement of human health. TR from bench to bedside involves a strong bidirectional relationship between basic science discovery and clinical practice. To support R&D planning and policy in HT effectively, the performance of TR programs was analyzed and evaluated in a R&D project on health and medical technology. TR programs were classified into three parts: unilateral TR, bilateral TR and multilateral TR. Bibliometrics and citation analysis were performed to assess research papers and gather information for the performance analysis of TR programs. In addition, both quantitative and qualitative analysis were successfully carried out using ISI Web of Science, Google Scholar Citations, SCOPUS and Knowledgematrix. In conclusion, the performance analysis of TR programs could significantly improve the efficiency of R&D plans, R&D management and evaluation for a safe and healthy life.
형질전환 벼 현탁세포 배양에서 hCTLA4Ig의 in situ 회수
최홍열(Hong-Yeol Choi),전수환(Su-Hwan Cheon),권준영(Jun-Young Kwon),윤보름(Boreum Yun),홍석미(Seok-Mi Hong),김선달(Sun-Dal Kim),김동일(Dong-Il Kim) 한국생물공학회 2016 KSBB Journal Vol.31 No.4
In this research, recombinant human cytotoxic Tlymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced by transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenic plant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promoter system which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell death promotes the release of protease which degrades the target proteins. The protein stability and productivity can be significantly influenced by proteolysis activity. Therefore, development of new strategies are necessary for the in situ recovery of target proteins from cell culture media. In this study, in situ recovery was performed by various strategies. Direct addition of Protein A resin with nylon bag leads to cell death by increased shear stress and decrease in production of hCTLA4Ig by protease. Medium exchange through modified flask could recover hCTLA4Ig with high cell viability and low protease activity, on the other hand, the productivity was lower than that of control. When in situ recovery was conducted at day 7 after induction in air-lift bioreactor, 1.94-fold of hCTLA4Ig could be recovered compared to control culture without in situ recovery. Consequently, in situ recovery of hCTLA4Ig from transgenic rice cell culture could enhance productivity significantly and prevent degradation of target proteins effectively.
형질전환 벼 현탁세포 배양에서 투과성 증진을 통한 hCTLA4Ig의 생산성 증대
최홍열(Hong-Yeol Choi),전수환(Su-Hwan Cheon),권준영(Jun-Young Kwon),임정애( Jung-Ae Lim),박혜림(Hye-Rim Park),김동일(Dong-Il Kim) 한국생물공학회 2016 KSBB Journal Vol.31 No.4
In this system, rice cells were genetically modified to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter induced by sugar depletion. Even though the target protein fused with signal sequence peptide, plant cell wall can be a barrier against secretion of recombinant proteins. Therefore, hCTLA4Ig can be trapped inside cell wall or remained in intracellular space. In this study, to enhance the secretion of hCTLA4Ig from cytoplasm and cell walls into the medium, permeabilizing agents, such as dimethyl sulfoxide (DMSO), Triton X-100 and Tween 20, were applied in transgenic rice cell cultures. When 0.5% (v/v) of DMSO was added in sugar-free medium, intracellullar hCTLA4Ig was increased, on the other hand, the secreted extracellular hCTLA4Ig was lower than that of control. DMSO did not give permeable effects on transgenic rice cell cultures. And Triton X-100 was toxic to rice cells and also did not give enhancing permeability of cells. When 0.05% (v/v) Tween 20 was added in rice cell cultures, however, intracellular hCTLA-4Ig was lower than that of control cultures. And the maximum 44.76 mg/L hCTLA4Ig was produced for 10 days after induction, which was 1.4-fold increase compared to that of control cultures. Especially, Tween 20 at 0.05% (v/v) showed the positive effect on the secretion of hCTLA4Ig though the decrease of intracellular hCTLA4Ig. Also, Tween 20 as a non-toxic surfactant did not affect the cell growth, cell viability and protease activity. In conclusion, secretion of hCTLA4Ig could be increased by enhancing permeability of cells regardless of the cell growth, cell viability and protease activity.
형질전환 벼 현탁세포를 이용한 hCTLA4Ig 생산에서 proline과 gelatin이 미치는 영향
송미나(Mi-Na Song),전수환(Su-Hwan Cheon),권준영(Jun-Young Kwon),최성훈(Sung-Hun Choi),김동일(Dong-Il Kim) 한국생물공학회 2009 KSBB Journal Vol.24 No.3
본 연구에서는 형질전환된 벼 세포를 이용하여 자가면역 질환의 치료제인 human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig)을 생산하였고, RAmy3D promoter와 RAmy1A signal peptide를 사용하여 당 고갈시에 목적 단백질의 발현과 배지로의 분비를 유도하였다. 이 시스템의 문제점은 당 고갈에 의해 목적 단백질의 생산이 유도되기 때문에 에너지원이 없어 세포의 사멸, 이로인한 배지내로의 protease 분비를 유발하게 된다. 이것은 목적 단백질의 안정성 저해와 궁극적으로는 생산성의 손실을 일으킨다. 따라서 세포를 보호하여 사멸을 막는 효과가 있음이 보고된 proline을 첨가하여 생산성 증진 효과를 확인하고자 하였다. 4 mM의 proline을 첨가하여 배양하였을 때, 세포의 사멸 및 protease의 분비를 줄일 수 있었고 이는 결과적으로 hCTLA4Ig의 생산성 증진으로 이어졌다. 또한 단백질 안정제를 통하여 배지내로 분비된 hCTLA4Ig 의 안정화를 이루어 생산성을 높이고자 하였다. 0.01 g/L 의 gelatin을 적용하여 생산성 증진 효과를 확인하였으며, 이는 hCTLA4Ig의 안정화에 의한 것임을 알 수 있었다. Protease의 분비를 줄이기 위한 세포 보호와 protease의 공격을 막기 위한 hCTLA4Ig 보호를 통하여 형질전환된 벼 세포를 이용한 hCTLA4Ig의 안정성 및 생산성을 증대 시킬 수 있음을 확인하였다. Rice cells were transformed to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter. hCTLA4Ig was produced and secreted into culture media inducibly when sugar was depleted. The obstacles of this system are the cell death and release of proteases by sugar starvation. These problems resulted in the losses of stability and productivity of hCTLA4Ig. Therefore, the effects of proline as an inhibitor of cell death were investigated. When 4 mM proline was added in sugar-free media, the cell death and release of proteases were reduced. As a consequence, the production of hCTLA4Ig was enhanced. In addition, the effects of protein stabilizers such as gelling agents were studied. It was found that the application of 0.01 g/L gelatin led to an increase in hCTLA4Ig production. This increase might be originated from the stabilization of hCTLA4Ig. In conclusion, the production of hCTLA4Ig could be enhanced by the additions of proline and gelatin in transgenic rice cell cultures.