표제지
요약
Abstract
목차
1. 서론 13
1.1. 바이오의약품 13
1.1.1. 바이오의약품 13
1.1.2. 바이오시밀러 13
1.2. 바이오의약품 시장 동향 15
1.3. Shake flask 17
1.3.1. Surface aeration 18
1.3.2. OUR 측정 방법 19
1.4. 산소 제한 (oxygen limitation) 20
2. 연구 목적 22
3. 재료 및 방법 23
3.1. Sodium sulfite oxidation 23
3.2. OTRmax 회귀분석[이미지참조] 25
3.3. 세포 배양 25
3.4. 배양액 분석 26
3.5. Actipro media DO 측정 26
4. 결과 및 고찰 27
4.1. 배양 환경에 따른 maximum viable cell density (VCDmax) 예측[이미지참조] 27
4.1.1. Sodium sulfite oxidation을 통한 배양 환경에 따른 OTRmax 측정[이미지참조] 27
4.1.2. 회귀분석을 통한 배양 환경에 따른 OTRmax 예측 모델 구축[이미지참조] 30
4.1.3. 배양 환경에 따른 Actipro 배지의 OTRmax 예측 모델 구축[이미지참조] 37
4.1.4. 배양 환경에 따른 maximum viable cell density (VCDmax) 예측[이미지참조] 37
4.2. CHO 세포 성장 및 대사에 대한 배양 환경의 OTRmax의 영향[이미지참조] 41
4.2.1. 배양 환경의 낮은 OTRmax에 의한 세포 성장 저해 확인[이미지참조] 41
4.2.2. 배양 환경의 낮은 OTRmax에 의한 세포 대사 변화 확인[이미지참조] 45
4.3. CHO 세포 배양 중 DO 변화 50
5. 결론 53
7. 참고문헌 55
Table 1. Classification of biopharmaceuticals 14
Table 2. Correlation of OTRmax to shaker rotation speed(n) and filling volume(VL) found in literature and rearranged for OTRmax [이미지참조] 34
Table 3. Literature values for CHO cell sOUR (sOUR : 10-10 mmol O2/cell*h) [이미지참조] 39
Table 4. VCDmax prediction by OTRmax calculation equation and CHO cell sOUR (OTRmax : mmol O2/L*h VCDmax : 107 cells/mL)[이미지참조] 40
Table 5. Comparison of lactate production rate (PL : 10-9 mg/cells*day)[이미지참조] 49
Table 6. Measured DO and sOUR (DO : %, sOUR : 10-10 mmol O₂/cellsxh)[이미지참조] 52
Figure 1. Global pharmaceutical market sales (A) and biopharmaceutical sales ratio (B) 16
Figure 2. Shake flask shape 17
Figure 3. Surface aeration 18
Figure 4. Occurrence of oxygen limitation 21
Figure 5. Effect of oxygen limitation on cell culture 21
Figure 6. Volume of 0.1 M sodium thiosulfate solution used in titration with time 28
Figure 7. Maximum oxygen transfer rate with shaker rotation speed and filling volume (5, 10, 15, 20, 25 mmol O₂/Lxh). 29
Figure 8. Result of regression equation 1 (A) and regression equation 2 (B) 33
Figure 9. Maximum oxygen transfer rate with shaker rotation speed (15, 20, 25, 30, 35, 40, 50, 60 mL). 35
Figure 10. Maximum oxygen transfer rate with filling volume (100, 130, 150, 200, 250 rpm). 36
Figure 11. Effect of OTRmax on VCD (A, C, E) and Viability (B, D, F). 100 rpm 25 mL (●), 100 rpm 50 mL (■), 150 rpm 25 mL (●), 150 rpm 50 mL (■), 200 rpm 25 mL (●), 200 rpm...[이미지참조] 44
Figure 12. Effect of OTRmax on lactate concentration (A, C, E) and lactate production rate (B, D, F) at 0.3, 1.5, 3x106 cells/mL (A), (B), (C). 100 rpm 25 mL (●), 100 rpm 50 mL (■), 150...[이미지참조] 48
Figure 13. Effect of OTRmax on DO at 0.3, 3x106 cells/mL (A), (B). (VCD : 100 rpm 25 mL (●), 100 rpm 50 mL (■), DO : 100 rpm 25 mL (□), 100 rpm 50 mL (□))[이미지참조] 51