Scale-down models are indispensable and crucial tools for process understanding and continuous process improvement in product life-cycle management. In this study, a scale-down model representing commercial-scale cell culture process of adalimumab biosimilar HS016 was first developed based on constant power per volume (P/V) principle and then qualified by multivariate data analysis (MVDA) and equivalence test method. The trajectories of the bench-scale process lie in the middle of the control range of large-scale process, built by multivariate evolution model based on nutrients, metabolites, and process performance datasets. This indicates that the small-scale process performance is comparable with that of the full-scale process. The final product titer, integrated viable cell density (iVCD), viability, aggregates, acid peak content, total afucosylation level, and high mannose content recognized as key process attributes (KPAs) or critical quality attributes (CQAs) were equivalent across the scales upon comparison using equivalence test method. The qualified scale-down model was then used for process characterization using a definitive screening design (DSD) where five independent variables including pH, shifted temperature, inoculation seeding density, viable cell density (VCD) at first feeding, VCD at temperature shift were evaluated. Three quadratic polynomial models for final product titer, aggregation, and high mannose were then established using the DSD results. The design space was finally developed using a probability-based Monte Carlo simulation method and was verified with the operation setpoint and worst-case condition. The case study presented in this report shows a feasible roadmap for cell culture process characterization.
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The authors would like to thank Dr. Khursheed Anwer from Celsion Corporation for manuscript review.
This study was supported by National Science and Technology Major Projects for “Major New Drugs Innovation and Development” (Grant No. 2018ZX09736008) and Science Program of Zhejiang province, China (Grant No. 2017C03003).
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Nie, L., Gao, D., Jiang, H. et al. Development and Qualification of a Scale-Down Mammalian Cell Culture Model and Application in Design Space Development by Definitive Screening Design. AAPS PharmSciTech 20, 246 (2019). https://doi.org/10.1208/s12249-019-1451-7
- scale-down model
- cell culture
- multivariate data analysis
- equivalence test
- definitive screening design