Pharmaceutical Biotechnology Quality by Design-Driven Process Development of Cell Culture in Bioreactor for the Production of Foot-And-Mouth Veterinary Vaccine Xinran Li 1 , 2 , Xuerong Liu 3 , Rongbin Wang 4, 5 , Fanglan An 3 , Jianqi Nie 1 , 2 , Yunde Zhang 3 , Hadji Ahamada 4, 5 , Xiuxia Liu 2, 5 , Chunli Liu 2, 5 , Yu Deng 2, 5 , Yankun Yang 1 , 2, 4, * , Zhonghu Bai 2, 4, 5, * 1 The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China 2 National Engineering Laboratory for Cereal FermentationTechnology, Jiangnan University, Wuxi 214122, China 3 China Agricultural Veterinary Biological Science and technology Co., Ltd, Lanzhou 730046, China 4 The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China 5 Jiangsu Provincial Research Center for Bioactive Product Processing Technology, Jiangnan University, Wuxi 214122, China article info Article history: Received 3 December 2018 Revised 21 January 2019 Accepted 8 February 2019 Keywords: quality by design vaccine cell culture batch processing biotechnology abstract Quality by design (QbD) principle has been established as a guideline to emphasize the understanding of the relationship of product quality with process control. Vaccine product have characteristics of security and high efficiency, but it also has features such as complexity and rigorous regulatory for production. This case study describes an example of QbD-driven process development for manufacturing a veterinary vaccine produced with baby hamster kidney-21 cells. The study revealed that cell culture duration was the most significant factor affecting 50% tissue culture infectious doses (TCID 50 ) and antigenic titer, and the factors of culture temperature and pH at infection phase exhibited less effect. Culture temperature at infection phase was the only significant factor for total protein. Through the Monte Carlo simulation, the design spaces of process parameters were determined. Meanwhile, the excellent and robust performance in manufacturing scale (4000-L) validated the effectiveness of this strategy. A reliable and robust multivariate process parameter range, that is, design space, was identified by this systematic approach. Our investigation presents a successful case of QbD principle, which encourages other researchers to combine the methodology into other biopharmaceutical manufacturing process. © 2019 American Pharmacists Association ® . Published by Elsevier Inc. All rights reserved. Introduction Quality by design (QbD) has been put forward to emphasize the importance of process control and product quality by the Interna- tional Conference on Harmonization (ICH). It has been adopted by the US Food and Drug Administration. The principles of pharma- ceutical development were outlined by the ICH Q8 (R2). 1 The annex of the ICH Q8 (R2) guidance document described the concepts and tools, which were used in pharmaceutical development to enhance products and improve process understanding. The ICH Q9 focused on quality risk management throughout product design and process development, which was essential for the enhanced development approach. 2 ICH Q10 contained elements of the phar- maceutical quality system and outlined the importance of product lifecycle management. 3 The ICH Q11 showed the method of applying the principle to the development and manufacturing of drug substance. 4 It further illustrated the concepts and tools described in ICH Q8 (R2) and quality risk management. Based on the improvement of methodology, the QbD approach intended to emphasize quality of product as an integral part of the develop- ment process. 5-7 This objective is achieved by establishing devel- opment targets upfront, scientific and risk-based assessment, statistical design of experiments, performing multifactorial explo- ration of the process design space, and establishing appropriate control strategies. So far to our best knowledge, there is no report mentioning on QbD principles application in foot-and-mouth dis- ease (FMD) vaccines production at large-scale using over 4000-L of bioreactors. The authors Xinran Li and Xuerong Liu contributed equally. * Correspondence to: Yankun Yang (Telephone: þ86 051085329306) and Zhonghu Bai (Telephone: þ86 051085197983). E-mail addresses: yangyankun@jiangnan.edu.cn (Y. Yang), baizhonghu@ jiangnan.edu.cn (Z. Bai). Contents lists available at ScienceDirect Journal of Pharmaceutical Sciences journal homepage: www.jpharmsci.org https://doi.org/10.1016/j.xphs.2019.02.004 0022-3549/© 2019 American Pharmacists Association ® . Published by Elsevier Inc. All rights reserved. Journal of Pharmaceutical Sciences xxx (2019) 1-8