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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2018, Vol. 38 Issue (6): 63-69    DOI: 10.13523/j.cb.20180609
    
Study on the Optimal Scale-down Model for Cell Growth and Mab-A Production of Sp2/0 Cells
Guo-qiang WANG1,2,*,Jian-ping LIU1,Hang ZHOU2,**()
1 School of Life Sciences Fudan University, Shanghai 200438, China
2 WuXi Biologics (Shanghai) Co., Ltd., Shanghai 200131, China
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Abstract  

Five different kinds of basal media were screened in batch mode to find out the optimal basal medium for cell growth of Sp2/0. The peak viable cell density in batch mode was 13.12×10 6cells/ml, and the culture duration was 7 days. Various cell culture conditions including different shaking speeds, concentrations of carbon dioxide, glutamine replaced by GlutaMAX TM, addition of trace element and different culture temperatures were studied in batch mode as well. Few different cell growth were found with these conditions except for culture temperatures, which leaded to different peak viable cell concentrations, different viabilites and then different culture durations. Fourteen kinds of combinations of feed media were screened in fed-batch mode. The peak viable cell densities were up to 30×10 6cells/ml, the culture duration were around 9 days, and the highest daily Mab-A production was 27.20mg/L. Batch re-feed mode was used in the third part of the study. The peak viable cell density was 50.42×10 6cells/ml, the culture duration was 14 days, and the highest daily Mab-A production (141.10mg/L) was 5.19 fold greater than in fed-batch mode. These studies suggest batch re-feed mode is the optimal scale-down mode for cell growth and Mab-A production of Sp2/0 cells.



Key words： Sp2/0      Scale-down mode      Batch re-feed     
Received: 24 December 2017      Published: 06 July 2018
ZTFLH:  Q256  
Corresponding Authors: Guo-qiang WANG,Hang ZHOU     E-mail: zhou_hang@wuxiapptec.com
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Guo-qiang WANG
Jian-ping LIU
Hang ZHOU
Cite this article:

Guo-qiang WANG,Jian-ping LIU,Hang ZHOU. Study on the Optimal Scale-down Model for Cell Growth and Mab-A Production of Sp2/0 Cells. China Biotechnology, 2018, 38(6): 63-69.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20180609     OR     https://manu60.magtech.com.cn/biotech/Y2018/V38/I6/63

Medium code Vendor Category
A,B ThermoFisher Basal medium
C,D,E GE
F,G ThermoFisher Feed medium
H,I,J,K GE
L Merck
M Kerry
N,O Lonza
P,Q Kerry Hydrolysate
R Merck Additive
Dexamethasone Merck
GlutaMAXTM ThermoFisher
Table 1 Medium information
Culture No. Medium Code Condition
SF1,5 A+C(50%+50%) Control
SF2,6 D
SF3,7 B
SF4,8 E
SF9 A+C(50%+50%) 160 RPM
SF10 D
SF11 B
SF12 E
SF13 A+C(50%+50%) 10% CO2
SF14 D
SF15 B
SF16 E
SF17 A+C(50%+50%) 33℃
SF19 E
SF18 A+C(50%+50%) 35℃
SF20 E
SF21 A+C(50%+50%) Trace element
SF22 E
SF23 A+C(50%+50%) GlutaMAXTM
SF24 D
SF25 B
SF26 E
Table 2 Conditions for batch culture
Culture No. Medium Code Conditon
SF27 99% F+1% L Feed medium screen
SF28 99% G+1% L
SF29 49.5% F+49.5% G+1% L
SF30 99% H+1% L
SF31 49.5% H+49.5% I+1% L
SF32 99% J+1% L
SF33 80% J+1% L+19% water
SF34 99% K+1% L
SF35 99% J (with 15 g/L P)+1% L
SF36 99% J (with 15 g/L Q)+1% L
SF37 99% N+1% L
SF38 99% M+1% L
SF39 99% O+1% L
SF40 99% J+1% L 36.5℃ shift to
33℃ on day 4
Table 3 Conditions for fed-batch culture
Fig.1 Cell growth (a) and viability (b) in various basal media
SF9 SF10 SF11 SF12 SF13 SF14 SF15 SF16 SF21 SF22 SF23 SF24 SF25 SF26
Peak VCC (106 cells/ml) 7.81 12.44 3.56 12.69 8.50 12.39 4.02 9.90 6.71 12.45 8.77 13.44 3.46 12.85
Culture duration (days) 5 7 4 7 5 7 4 7 5 7 5 7 4 7
Table 4 Influence of shaking speed, CO2, trace element and GlutaMAXTM on cells’ performance
Fig.2 Cell growth (a) and viability (b) under different culture temperatures
Culture
No.		 Peak VCC
(106
cells/ml)		 Culture
duration
(days)		 Titer
(mg/L)		 qMab-A,
(pg/cell/
day)
SF27 25.65 7 73.73 0.99
SF28 27.56 7 35.40 0.46
SF29 26.57 7 46.93 0.63
SF30 20.58 7 39.69 0.68
SF31 21.49 7 41.39 0.74
SF32 23.26 7 138.58 1.97
SF33 20.48 8 99.14 1.28
SF34 22.96 6 41.74 0.87
SF35 37.08 9 165.92 1.23
SF36 20.29 8 157.20 1.83
SF37 25.63 6 42.94 0.83
SF38 22.57 6 47.85 1.02
SF39 22.91 6 56.72 1.13
SF40 20.52 8 217.59 2.48
Table 5 Cells’ performance in various feed media
Culture
No.		 Peak VCC
(106
cells/ml)		 Culture
duration
(days)		 Titer
(mg/L)1)		 qMab-A,
(pg/cell/
day)
ST1 50.42 14 141.10 2.98
ST2 42.23 14 115.38 2.77
ST3 53.01 14 162.56 3.19
Table 6 Cells’ performance in batch re-feed mode
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