Ambr™ Mini-bioreactor as a high-throughput tool for culture process development to accelerate transfer to stainless steel manufacturing scale: comparability study from process performance to product quality attributes

Abstract

Background Enhancing throughput of bioprocess development has become increasingly important to rapidly screen and optimize cell culture process parameters. With increasing timeline pressures to get therapeutic candidates into the clinic, resource intensive approaches such as the use of shake flasks and bench-top bioreactors may limit the design space for experimentation to yield highly productive processes. The need to conduct large numbers of experiments has resulted in the use of miniaturized high-throughput (HT) technology for bioprocess development. One such high-throughput system is the ambrTM platform, a robotically driven, mini-bioreactor system developed by TAP-Sartorius. In this study we assessed and compared the performance parameters of ambrTM mini-bioreactor runs to 2L glasses , 80L and 400L stainless steel bioreactors using a CHO cell line producing a recombinant monoclonal antibody. The daily parameters monitored during the cultures were cell growth and cell viability, offline pH and dissolved oxygen, metabolite profiles (glucose, lactate and ammonia) and monoclonal antibody titer. In addition, we compared the product quality attributes (high and low molecular weight species, charge variants) of the clarified cell culture fluid post Protein-A elution generated in the mini-bioreactor run to the larger manufacturing scales. Materials and methods A genetically engineered Dihydrofolate Reductase (DHFR)-/ DG44 Chinese Hamster Ovary (CHO) cell line with Methotrexate (MTX) as a selective agent, expressing a recombinant monoclonal antibody was used. Cells were cultivated for 14 days in a fed-batch mode in a chemically defined medium and fed according to process description. Culture systems: Different bioreactor scales were used in this study : ambrTM48 (TAP -Sartorius Biosystems), an automated system with 48 disposable microbioreactor vessels, 2L stirred tank glasses bioreactors with Biostat B-DCUII control systems (Sartorius Stedim), 80L and 400L stainless-steel bioreactors (Zeta). Data was analysed using JMP statistical (SAS) program. All the experiments were conducted using the same cell bank at the same cell age at bioreactor inoculation. pH (7.0 +/0.2) was controlled using CO2 and base addition. The scale independent factors (pH, DO set point, seeding density, temperature, culture duration, media and feed composition), were the same for all the scales. The scale dependent factors (culture start volume, feed volumes) were linearly adapted. Agitation speed and aeration that were determined theoretically or though experiment. Sampling plans and sample volumes were especially adapted to the ambrTM system to take into account the low bioreactor volume. * Correspondence: frederic.delouvroy@ucb.com Upstream Process Sciences, Biotech Sciences, UCB Pharma S.A., Chemin du Foriest, Braine l’Alleud, Belgium Delouvroy et al. BMC Proceedings 2015, 9(Suppl 9):P78 http://www.biomedcentral.com/1753-6561/9/S9/P78