원문정보
Enhanced Production of hCTLA4Ig by Adding Sodium Butyrate and Sodium Pyruvate
초록
영어
Human cytotoxic T-lymphocyte antigen 4- immunoglobulin (hCTLA4Ig), an immunosuppressive agent, was expressed in transgenic rice cells using RAmy3D promoter and RAmy1A signal peptide for the inducible production and secretion into culture media by sugar depletion. In this study, sodium butyrate was used as a small molecular enhancer (SME) to enhance the production of hCTLA4Ig in transgenic rice cell suspension cultures. When 1 mM sodium butyrate was added in sugar-free media, relative viability was not reduced, while the productivity was improved 1.3-fold. In addition, by supplementing 87 mM sodium pyruvate as an alternative energy source during the production phase, death rate of the cells was decreased. When sodium pyruvate was not added, most cells became dead at day 6. However, by adding sodium pyruvate, 18% of viability can be maintained until day 10 and the production of hCTLA4Ig was enhanced 1.4-fold. When the combination of sodium pyruvate and sodium butyrate at optimum concentrations was added, the highest viability and hCTLA4Ig production could be obtained. The highest level of hCTLA4Ig reached up to 35 mg/L at day 10.
목차
1. 서론
2. 재료 및 방법
2.1. 세포주 및 세포배양
2.2. 세포량 측정
2.3. Cell viability 측정
2.4. Protease 활성 측정
2.5. 총단백질 정량
2.6. hCTLA4Ig 정량
2.7. Total genomic DNA 분석
3. 결과 및 고찰
3.1. Sodium butyrate 첨가가 미치는 영향
3.2. 에너지원 첨가의 영향
3.3. Sodium butyrate와 sodium pyruvate 동시 첨가에 따른 영향
4. 결론
감사
References
