표제지
요약
Abstract
목차
1. 서론 13
1.1. 동물세포배양 13
1.1.1. Chinese Hamster ovary cell 13
1.2. 동물세포배양배지 16
1.2.1. 동물세포배양배지 종류 16
1.2.2. 무 혈청배지 구성요소 18
1.3. Design of experiments (DoE) 25
1.3.1. Plackett-burman design (PBD) 27
1.3.2. Fractional factorial design 28
1.3.3. Central composite design (CCD) 30
2. 연구 목적 32
3. 재료 및 방법 34
3.1. 세포배양 34
3.2. 배지제조 35
3.3. Viable cell density, cell viability 측정 38
3.4. Design of experiments (DoE) 38
3.4.1. Plackett-burman design (PBD) 40
3.4.2. Fractional factorial design 42
3.4.3. Central composite design (CCD) 43
4. 결과 44
4.1. ICP-MS analysis 44
4.2. Amino acid analysis 44
4.3. Plackett-burman design (PBD) 48
4.4. Fractional factorial design 52
4.5. Central composite design (CCD) 55
5. 결론 58
6. 참고문헌 61
Table 1. Natural and artificial media used in animal cell culture. 18
Table 2. Essential amino acids and non-essential amino acids. 23
Table 3. Organize terms used in Design of experiences (DoE) 26
Table 4. The number and level of factor and experiments of a Fractional factorial design. If the level is n and the number of... 29
Table 5. List of components used in the manufacture of medium. 37
Table 6. The Plackett-burman design (PBD) table for 27 factors and 4 center point. 41
Table 7. The Fractional factorial design table for 27 factors and 3 center point. 42
Table 8. The Central composite design (CCD) table for 3 factors. 43
Figure 1. Types and history of CHO cells. Dotted lines indicate uncertainty and small arrows indicate distribution in... 15
Figure 2. The mechanism for controlling pH from medium to CO₂. The two reactions seek equilibrium. Therefore, When... 24
Figure 3. Design of experiments (DoE) stage. There are three stages: screening stage, characterization stage and optimization stage. 26
Figure 4. (a) (b) A comparison between a fractional factorial design and a full factor design for a two-level design with... 29
Figure 5. Design point of CCD. (a) The black dot is Factorial Cube points, (b) The green dot is axial points. It is located... 31
Figure 6. Type of Central composite design (CCD) axis point. (a) circumscribed center design (b) inscribed center design.... 31
Figure 7. Medium development scheme. seven factors were selected at the screening DoE stage, three factors were selected at the Characterization DoE stage, and serum-free medium was... 39
Figure 8. Results of ICP-MS Analysis of Three Commercialized Media. Three Commercialized media were... 46
Figure 9. Results of Amino acid analysis of Three Commercialized Media. Three Commercialized media were... 47
Figure 10. Results of Peak viable cell density in Plackett-burman design (PBD) 49
Figure 11. Results of pareto chart in Plackett-burman design (PBD) 50
Figure 12. Results of main effect chart in Plackett-burman design (PBD) 51
Figure 13. Results of Peak viable cell density in Fractional factorial design. 53
Figure 14. Results of pareto chart in Fractional factorial design. 53
Figure 15. Results of main effect chart in Fractional factorial design 54
Figure 16. Regression equation from Central Composite Design (CCD) to PVCD value. 56
Figure 17. Results of Peak viable cell density in Central composite design (CCD). 56
Figure 18. Results of contour chart in Central composite design (CCD). 57
Figure 19. Day 4 Peak Viable Cell Density Comparison Chart. 60