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

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.

Perfusion Processes

“…Applications of perfusion mode other than for the manufacturing of biopharmaceuticals have recently been reported and are increasingly used in the industry, such as cell bank manufacturing (Tao et al 2011;Seth et al 2013;Clincke et al 2013a) or high cell density seeding of production bioreactors, i.e. 'N-1 step', (Pohlscheidt et al 2013;Padawer et al 2013;Yang et al 2014;Hecht et al 2014).…”

Development of small scale cell culture models for screening poloxamer 188 lot‐to‐lot variation

“…The rising demand of therapeutic monoclonal antibodies and recombinant protein products boosted the US biotechnology sales over 60 billion USD in 2012 . In order to meet the increasing manufacturing output required to satisfy commercial and clinical demands, cell culture technology has made significant progress in increasing productivity over the past several decades, which include engineering of cell lines, optimization of culture media, improvement of feed strategy, and modification of operational parameters …”

“…1 In order to meet the increasing manufacturing output required to satisfy commercial and clinical demands, cell culture technology has made significant progress in increasing productivity over the past several decades, which include engineering of cell lines, 2,3 optimization of culture media, improvement of feed strategy, [4][5][6] and modification of operational parameters. [7][8][9] In recent years, the lot to lot variability of raw materials has received significant attention in drug manufacturing. Culture media is composed of more than 50 chemical or biological compounds, which often makes it difficult to identify the root cause of process variation.…”

Bioreactor concepts for cell culture-based viral vaccine production

“…Second, perfusion systems can be used feeding fresh medium, while withdrawing spent medium to achieve higher cell concentrations and extended run times beyond typical upper limits of batch and fed-batch processes. This benefit gives the option to operate a single perfusion bioreactor as constant seed reactor for multiple batch processes in parallel or even time-shifted (hybrid processing) [120]. These cell separation devices mainly work in cross flow mode with either constant, pulsed or alternating flow directions avoiding membrane fouling and promoting disaggregation of cell clumps due to increased shear forces.…”