형질전환 벼 현탁세포 배양에서 hCTLA4Ig의 in situ 회수In situ Recovery of hCTLA4Ig from Suspension Cell Cultures of Oryza sativa
- Other Titles
- In situ Recovery of hCTLA4Ig from Suspension Cell Cultures of Oryza sativa
- Authors
- 최홍열; 전수환; 권준영; 윤보름; 홍석미; 김선달; 김동일
- Issue Date
- 2016
- Publisher
- 한국생물공학회
- Keywords
- Plant cell culture; In situ recovery; hCTLA4Ig; Airlift bioreactor; Protease
- Citation
- KSBB Journal, v.31, no.4, pp.284 - 290
- Journal Title
- KSBB Journal
- Volume
- 31
- Number
- 4
- Start Page
- 284
- End Page
- 290
- URI
- https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/8890
- DOI
- 10.7841/ksbbj.2016.31.4.284
- ISSN
- 1225-7117
- Abstract
- In this research, recombinant human cytotoxic Tlymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was producedby transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenicplant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promotersystem which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell deathpromotes 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.
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