원문정보
Enhanced Production of hCTLA4Ig by Suppressing Cell Death in Transgenic Rice Cell Suspension Cultures
초록
영어
Transgenic plant cell cultures are an attractive expression system for the production of industrial and pharmaceutical proteins because of their advantages in safety and low production cost. Human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced and secreted when sugar was depleted in culture medium by transgenic rice cell lines (Oryza sativa L.) using RAmy3D promoter. Due to the production of the target protein by sugar depletion, concomitant occurrence of cell death is inevitable. For that reason, inhibition of cell death for enhancing productivity was necessary for the production period without energy sources. Supplementation of 0.1 mM sodium nitroprusside improved cell viability by 1.4-fold and maximum hCTLA4Ig production by 1.3-fold compared to those of control. Addition of 1 and 10 mM glutathione, N-acetylcysteine (NAC), and nicotinamide inhibited apoptotic-like programmed cell death by decreasing the activity of reactive oxygen species. Production hCTLA4Ig was enhanced 1.4-, 1.25-, and 1.15-fold with 10 mM NAC, 1 mM NAC, and 1 mM glutathione, respectively. In addition, it was found that the supplementation of NAC enhanced the cell viability.
목차
1. 서론
2. 재료 및 방법
2.1. 세포주 및 배양 조건
2.2. 세포량 측정
2.3. 세포 생존도 측정
2.4. hCTLA4Ig의 정량분석
2.5. Protease 활성 측정
2.6. DNA fragmentation 분석
2.7. Autophagosome 형성 확인
3. 결과 및 고찰
3.1. 세포사멸 확인
3.2. Sodium nitroprusside의 영향
3.3. 항산화제 첨가의 효과
4. 결론
감사
REFERENCES