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Production of foreign proteins by transgenic plant cell cultures has several advantages such as post-translational modification, low risk of product contamination and low-cost production and purification. However, target proteins are degraded by extracellular proteases existing in the media. A solution to this problem is the use of perfusion culture and ion exchange chromatography for the application of integrated bioprocess using in situ recovery. With this method, production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was investigated in this study. First, optimization of cell concentration during the induction phase for the production of hGM-CSF was examined. As cell concentration increased, the level of hGM-CSF was decreased due to the presence of extracellular proteases. Induction using sugarfree media produced 33% more hGM-CSF. The effects of pH on the binding of hGM-CSF to cationic and anionic exchange resins were also investigated. In terms of stability, optimal pH was found to be 5~7. In the case of using buffer exchange when CM-Sepharose was used as a cationic exchange resin, optimal pH for binding was 4.8 and adsorption yield was 77%.

When DEAE-Sepharose was used as an anionic exchange resin, it was 5.5 (74%). Without buffer exchange, optimal pH was 4.6 (84%). From these results, an integrated bioprocess using in situ recovery with simultaneous production and separation of foreign protein in transgenic plant cell suspension cultures was found to be feasible.

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Desulfovibrio Desulfuricans과 제올라이트를 이용한 해양 내의 Zn, As 제거용 미생물 담체 개발 김인화, 최진하, 주정옥, 오병근 pp.114-118
염 환경 하에서 Bacillus sp. SH1RP8와 Polyacrylate Polymers가 작물생장에 미치는 영향에 관한 연구 홍선화, 김지슬, 박장우, 이은영 pp.97-102
형질전환 벼 현탁세포 배양에서 hGM-CSF의 in situ Recovery 연구 명현종, 최홍열, 남형진, 김동일 pp.103-108
프로바이오틱스의 동결보호 및 장관안정성 개선을 위한 첨가제 효과분석 정은지, 문대원, 오준석, 문진석, 김광엽, 최혜선 pp.109-113
항산화능이 증대된 Arthrospira platensis 균주 개량 및 특성 분석 최수정, 이재화 pp.119-124
The Application of Thermotolerant Yeast Kluyveromyces marxianus as a Potential Industrial Workhorse for Biofuel Production 박재범, 김진성, 장승원, 홍은수, 하석진 pp.125-131
곤충병원성 선충에서 분리한 공생세균의 안정화 및 항진균활성 강동희, 김효현, 남욱호, 김현수 pp.132-139

참고문헌 (19건) : 자료제공( 네이버학술정보 )

참고문헌 목록에 대한 테이블로 번호, 참고문헌, 국회도서관 소장유무로 구성되어 있습니다.
번호 참고문헌 국회도서관 소장유무
1 James, E. A., C. Wang, Z. Wang, R. Reeves, J. H. Shin, N. S. Magnuson, and J. M. Lee (2000) Production and characterization of biologically active human GM-CSF secreted by genetically modified plant cells. Protein Expr. Purif. 19: 131-138. 미소장
2 Zhang, X. W., T. Sun, X. Liu, D. X. Gu, and Z. Q. Tang (1999)Production of granulocyte macrophage colony stimulating factor (GM-CSF) by high cell density fermentation of secretory recombination Escherichia coli. Process Biochem. 34: 55-58. 미소장
3 Biotechnology: Promethean Science or Obsession? 네이버 미소장
4 Biotechnology in Turkey: an overview. 네이버 미소장
5 Twyman, R. M., E. Stoger, S. Schillberg, P. Christou, and R. Fischer (2003) Molecular farming in plants: Host systems and expression technology. Trends Biotechnol. 21: 570-578. 미소장
6 Mascia, P. N. and R. B. Flavell (2004) Safe and acceptable strategies for producing foreign molecules in plants. Curr. Opin. Plant. Biol. 7: 189-195. 미소장
7 Grabowski, G. A., M. Golembo, and Y. Shaaltiel (2014) Taliglucerase alfa: An enzyme replacement therapy using plant cell expression technology. Mol. Gen. Metabol. 112: 1-8. 미소장
8 Magnuson, N. S., P. M. Linzmaier, J.-W. Gao, R. Reeves, G. An, and J. M. Lee (1996) Enhanced recovery of secreted mammalian protein from suspension culture of genetically modified tobacco cells. Protein Expr. Purif. 7: 220-228. 미소장
9 Hooker, B. S. and J. M. Lee (1990) Cultivation of plant cells in aqueous two-phase polymer systems. Plant Cell Rep. 8: 546-549. 미소장
10 Biotechnology Regulation in Europe 네이버 미소장
11 James, E., D. R. Mills, and J. M. Lee (2002) Increased production and recovery of secreted foreign proteins from plant cell cultures using an affinity chromatography bioreactor. Biochem. Eng. J. 12:205-213. 미소장
12 Terashima, M., N. Hashikawa, M. Hattori, and H. Yoshida (2002)Growth characteristics of rice cell genetically modified for recombinant human a1-antitrypsin production. Biochem. Eng. J. 12: 155-160. 미소장
13 Kwon, J. Y., K. H. Lee, S. H. Cheon, and D. I. Kim (2012) Application of anoxia with glucose addition for the enhanced production of hCTLA4Ig in transgenic rice suspension cell cultures. Enzyme Microb. Technol. 50: 298-303. 미소장
14 James, E. A. and J. M. Lee (2001) The production of foreign protein from genetically modified plant cells. Adv. Biochem. Eng. Biotechnol. 72: 127-156. 미소장
15 Kusnadi, A. R., Z. L. Nikolov, and J. A. Howard (1997) Production of recombinant protein in transgenic plants: practical considerations. Biotechnol. Bioeng. 56: 473-484. 미소장
16 Magnuson, N. S., P. M. Linzmaier, R. Reeves, G. An, K. Hay-Glass, and J. M. Lee (1998) Secretion of biologically active human interleukin-2 and interleukin-4 from genetically modified tobacco cells in suspension culture. Protein Express. Purif. 13: 45-52. 미소장
17 Biotechnology in the Eastern Bloc 네이버 미소장
18 Biotechnology: Acquired or Consumed? 네이버 미소장
19 Choi, J. W., G. H. Cho, S. Y. Byun, and D. I. Kim (2001) Integrated bioprocessing for plant cell cultures. Adv. Biochem. Eng. Biotechnol. 72: 63-103. 미소장

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