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Molecular Farming Symposium
Process Development for the Production of hCTLA4Ig Using Transgenic Rice Cells in Bioreactor
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- 발행기관
- 한국생물공학회 바로가기
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- 간행물
- 한국생물공학회 학술대회 바로가기
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- 통권
- 2009 춘계학술대회 및 국제심포지움 (2009.04)바로가기
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- 페이지
- pp.76-76
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- 저자
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- 언어
- 영어(ENG)
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- URL
- https://www.earticle.net/Article/A104616
※ 원문제공기관과의 협약기간이 종료되어 열람이 제한될 수 있습니다.
원문정보
초록
- 영어
- Human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig), a fusion recombinant protein, was expressed in transgenic rice cell suspension cultures. The expression vector pMYN409 was constructed to express hCTLA4Ig under the control of RAmy3D promoter. Under the induction condition by sugar starvation, suspension-cultured rice cells secreted hCTLA4Ig into the media. The rice-derived hCTLA4Ig could be purified with affinity chromatography using protein A resin. Recombinant hCTLA4Ig has molecular weight of ~50 kDa on SDS-PAGE under
reducing condition. hCTLA4Ig was produced inducibly by sugar starvation in stirred tank bioreactors with various process strategies. Marine, turbine, and hollowed paddle impeller were compared for the cultivation of transgenic rice cells and optimization of culture medium was performed. Two-stage cultivation method was applied to achieve high cell density during cell growth phase and to enhance the production of hCTLA4Ig. Two kinds of media exchange methods (perfusion and direct exchange) were compared in a 5-L stirred tank bioreactor. In the production phase, inducible production of hCTLA4Ig caused cell death because of sugar depletion, and media exchange induced cell lysis by additional shear stress. It was essential to reduce shear stress for increasing productivity because protease activity was increased by cell lysis. High density cultivation was also investigated in the bioreactor. With fed-batch
cultivation, maximum dry cell weight reached 21.2 g/L and maximum hCTLA4Ig production reached 13.4 mg/L which were 2.18-fold and 2.14-fold higher than those of the direct media exchange, respectively. Therefore, we can conclude that the minimization of cell lysis and achievement of high cell density could be necessary to enhance the production of hCTLA4Ig with RAmy3D promoter system. Control of oxygen level was found to be important. In addition, effect of shear was significant. S225
저자
간행물 정보
발행기관
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- 발행기관명
- 한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
- 설립연도
- 1984
- 분야
- 공학>생물공학
- 소개
- 이 법인은 생물 공학의 발전과 보급에 이바지하고, 회원 상호 간의 연구 협력과 친목을 도모함을 목적으로 한다 1. 생물공학 분야의 발전을 위한 연구 협력 2. 생물공학의 실용화를 촉진시키기 위한 산학 협동 3. 학술연구 발표회, 강연회, 연수회 등 학술활동의 개최 4. 국,영문 학술지,소식지,학술회의 Proceedings 및 학술도서의 발간 5. 생물공학 발전을 위한 정책 건의 6. 기타 국제 교류 등 본 학회의 목적 달성을 위한 제반 활동
간행물
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- 간행물명
- 한국생물공학회 학술대회
- 간기
- 반년간
- 수록기간
- 1985~2013
- 십진분류
- KDC 476 DDC 576
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