Rational Design and Establishment of CHO Cell Intensified Fed-batch Culture Process

doi:10.13523/j.cb.2305046

China Biotechnology

2023, Vol. 43

Issue (10): 20-31 DOI: 10.13523/j.cb.2305046

Rational Design and Establishment of CHO Cell Intensified Fed-batch Culture Process

SU Ying¹,ZHANG Wei-jian¹,WAN Yu-xiang²,SHEN Tian-fang²,ZHANG Xin-ran¹,ZHANG Ru-yue¹,TAN Wen-song^1,³,ZHAO Liang^1,^3,^*(

)

1 State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China
2 Zhejiang Hisun Pharmaceutical Co., Ltd., Hangzhou 311404, China
3 Shanghai Bioengine Sci-Tech Co., Ltd., Shanghai 201203, China

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Abstract

Objective: Due to the higher seeding density and running density of the intensified fed-batch of ultra-high seeding density (uHSD-IFB) process, the feeding strategy of the traditional low-density culture process often does not provide sufficient nutrients for cell maintenance and product expression of the process, which ultimately leads to low process yield and lower production efficiency. By optimizing the feed medium and feeding strategy, the uHSD-IFB process will be successfully established and the target protein yield will be increased. Methods: A monoclonal antibody-expressing CHO-K1 cell line was studied in this paper, a two-stage dynamic feedback feeding strategy with glucose as the control indicator was designed by metabolic kinetics and stoichiometry analysis, and the nutrient concentrations for key trace elements in the feed medium were screened and optimized in combination with design of experiment (DoE). Results: The optimized process effectively alleviated the contradiction between nutrient depletion and accumulation of metabolic by-products in the uHSD-IFB process and achieved the purpose of cell growth and product synthesis in the ultra-high inoculation density culture process. The cumulative titer of the optimized design of the uHSD-IFB process was increased by 95% and the daily yield was increased by about 97% compared to that before optimization. Conclusion: This proposed feeding strategy can provide help to rapidly establish an enhanced fed-batch culture process of ultra-high seeding density with high cell density and high product expression.

Key words： CHO cell Process intensification Dynamic feedback fed-batch Rational design Glucose control model

Received: 30 May 2023 Published: 02 November 2023

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	SU Ying
	ZHANG Wei-jian
	WAN Yu-xiang
	SHEN Tian-fang
	ZHANG Xin-ran
	ZHANG Ru-yue
	TAN Wen-song
	ZHAO Liang

Cite this article:

SU Ying, ZHANG Wei-jian, WAN Yu-xiang, SHEN Tian-fang, ZHANG Xin-ran, ZHANG Ru-yue, TAN Wen-song, ZHAO Liang. Rational Design and Establishment of CHO Cell Intensified Fed-batch Culture Process. China Biotechnology, 2023, 43(10): 20-31.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2305046 OR https://manu60.magtech.com.cn/biotech/Y2023/V43/I10/20

Table 1 Glucose control model for amino acid concentrations in feed medium during uHSD-IFB process

Fig.1 Rational design workflow diagram for the optimized uHSD-IFB process

Fig.2 Comparison of cell growth and protein production in traditional and intensified fed-batch process (a) Viable cell density and viability (b) Titer and Vp

Fig.3 Concentration change of amino acids during uHSD-IFB process

Fig.4 Specific consumption (production) rate of amino acids during uHSD-IFB (AA) process

Fig.5 Relationship between cumulative consumption of amino acids and cumulative consumption of glucose in the cell growth phase and product production phase during uHSD-IFB (AA) process (a) Cell growth phase (b) Product production phase

Fig.6 Comparison of the uHSD-IFB process for the glucose control model with the uHSD-IFB process before optimization (a) Cell growth (b) Protein production (c) Glucose concentration (d) Lactate concentration

Table 2 Factors and levels of Plackett-Burman experiments design

Table 3 Variance analysis of Plackett-Burman experiments results

Table 4 Comparison of the main results of several fed-batch processes


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