Development of a new bioprocess scheme using frozen seed train intermediates to initiate CHO cell culture manufacturing campaigns

TLDR

Agility to schedule and execute cell culture manufacturing campaigns quickly in a multi‑product facility will play a key role in meeting the growing demand for therapeutic proteins. The study investigates whether initiating campaigns with cryopreserved CHO cells in large‑volume bags can shorten timelines, maximize plant flexibility, and reduce resource use, and reviews the optimization of scaling, freezing, and rack configuration steps. FASTEC cultivates CHO cells to high density in a perfusion bioreactor, cryopreserves them in disposable bags using a controlled‑rate protocol, thaws the bags for each campaign, and employs flow cytometry to assess post‑freezing recovery. Development data show that FASTEC produces drug with quantity and quality comparable to conventional seed‑train processes, thereby offering flexible manufacturing options for the cell‑culture network.

Abstract

Agility to schedule and execute cell culture manufacturing campaigns quickly in a multi-product facility will play a key role in meeting the growing demand for therapeutic proteins. In an effort to shorten campaign timelines, maximize plant flexibility and resource utilization, we investigated the initiation of cell culture manufacturing campaigns using CHO cells cryopreserved in large volume bags in place of the seed train process flows that are conventionally used in cell culture manufacturing. This approach, termed FASTEC (Frozen Accelerated Seed Train for Execution of a Campaign), involves cultivating cells to high density in a perfusion bioreactor, and cryopreserving cells in multiple disposable bags. Each run for a manufacturing campaign would then come from a thaw of one or more of these cryopreserved bags. This article reviews the development and optimization of individual steps of the FASTEC bioprocess scheme: scaling up cells to greater than 70 × 10(6) cells/mL and freezing in bags with an optimized controlled rate freezing protocol and using a customized rack configuration. Flow cytometry analysis was also employed to understand the recovery of CHO cells following cryopreservation. Extensive development data were gathered to ensure that the quantity and quality of the drug manufactured using the FASTEC bioprocess scheme was acceptable compared to the conventional seed train process flow. The result of offering comparable manufacturing options offers flexibility to the cell culture manufacturing network.