Development of an intensified fed-batch production platform with doubled titers using N-1 perfusion seed for cell culture manufacturing

Jianlin Xu,Michael C Borys,Charles Hill,Mengmeng Xu,Matthew S Rehmann,Shun Zheng,Zheng Jian Li,Qin He

doi:10.1186/s40643-020-00304-y

Jianlin Xu, Michael C Borys + Show 6 more

Open Access

https://doi.org/10.1186/s40643-020-00304-y

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Abstract

The goal of cell culture process intensification is to increase volumetric productivity, generally by increasing viable cell density (VCD), cell specific productivity or production bioreactor utilization in manufacturing. In our previous study, process intensification in fed-batch production with higher titer or shorter duration was demonstrated by increasing the inoculation seeding density (SD) from ~ 0.6 (Process A) to 3–6 × 106 cells/mL (Process B) in combination with media enrichment. In this study, we further increased SD to 10–20 × 106 cells/mL (Process C) using perfusion N-1 seed cultures, which increased titers already at industrially relevant levels by 100% in 10–14 day bioreactor durations for four different mAb-expressing CHO cell lines. Redesigned basal and feed media were critical for maintaining higher VCD and cell specific productivity during the entire production duration, while medium enrichment, feeding strategies and temperature shift optimization to accommodate high VCDs were also important. The intensified Process C was successfully scaled up in 500-L bioreactors for 3 of the 4 mAbs, and quality attributes were similar to the corresponding Process A or Process B at 1000-L scale. The fed-batch process intensification strategies developed in this study could be applied for manufacturing of other mAbs using CHO and other host cells.

Highlights

Monoclonal antibody product development has had great success with over 80 mAbs granted marketing approvals, while the commercial pipeline is robust with over 570 mAbs in different clinical development stages (Ecker et al 2015; Kaplon and Reichert 2019; Walsh 2018)
Great progress has been made in mAb process development, the manufacturing cost remains much higher for mAbs than small molecules
We present the development of an intensified fed-batch platform for production of multiple mAbs using Chinese hamster ovary (CHO) K1 GS cell lines

Summary

Introduction

Monoclonal antibody (mAb) product development has had great success with over 80 mAbs granted marketing approvals, while the commercial pipeline is robust with over 570 mAbs in different clinical development stages (Ecker et al 2015; Kaplon and Reichert 2019; Walsh 2018). Global annual sales of mAbs were above $115 billion in 2018 and are predicted to rise above $300 billion in 2025 (Lu et al 2020). These great achievements are partially due to the higher approval success rates for mAbs than small molecule drug products (Hay et al 2014; Kaplon and Reichert 2019). Great progress has been made in mAb process development, the manufacturing cost remains much higher for mAbs than small molecules. Higher titers continue to be reported in recent literature (e.g. 9–10 g/L titer in 14–18 days) (Handlogten et al 2018; Huang et al 2010; Takagi et al 2017), suggesting that potential remains for further increases in productivity and reduction in cost using fed-batch operation

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