Impact of media and antifoam selection on monoclonal antibody production and quality using a high throughput micro‐bioreactor system

Abstract

Monoclonal antibody production in commercial scale cell culture bioprocessing requires a thorough understanding of the engineering process and components used throughout manufacturing. It is important to identify high impact components early on during the lifecycle of a biotechnology derived product. While cell culture media selection is of obvious importance to the health and productivity of mammalian bioreactor operations, other components such as antifoam selection can also play an important role in bioreactor cell culture. Silicone polymer based antifoams were known to have negative impacts on cell health, production, and downstream filtration and purification operations. High throughput screening in micro-scale bioreactors provides an efficient strategy to identify initial operating parameters. Here, we utilized a micro-scale parallel bioreactor system to study an IgG1 producing CHO cell line, to screen Dynamis, ProCHO5, PowerCHO2, EX-Cell Advanced and OptiCHO media, and 204, C, EX-Cell, SE-15 and Y-30 antifoams and their impacts on IgG1 production, cell growth, aggregation, and process control. This study found ProCHO5, EX-Cell Advanced and PowerCHO2 media supported strong cellular growth profiles, with an IVCD of 25–35 × 106 cells-d/mL, while maintaining specific antibody production (Qp>2 pg/cell-d) for our model cell line and a monomer percentage above 94%. Antifoams C, EX-Cell and SE-15 were capable of providing adequate control of foaming while antifoam 204 and Y-30 noticeably stunted cellular growth. This work highlights the utility of high throughput micro bioreactors and the importance of identifying both positive and negative impacts of media and antifoam selection on a model IgG1 producing CHO cell line.

Real‐time quantification and supplementation of bioreactor amino acids to prolong culture time and maintain antibody product quality

“…This experiment used a previously described recombinant CHO DG44 cell line that expresses a model chimeric IgG1 . For all the reactors subjected to feed strategies 1 through 3, 5, and 6, the seed train was started by thawing 1 vial (1 ml) of banked cells (7 × 10 7 viable cells/ml) into 200 ml of 37°C CD OptiCHO media (Life Technologies, A11222) supplemented with 1X Penicillin/Streptomycin (Corning, 30‐001‐Cl) and 8 mM l ‐glutamine (Corning, 25‐005‐CV) in a 1 L spinner flask (Corning, 4500‐1L).…”

Impacts of intentional mycoplasma contamination on CHO cell bioreactor cultures

“…This experiment used a previously described recombinant CHO DG44 cell line that expresses a model chimeric IgG1 (Velugula‐Yellela, Williams, et al, ). Frozen cell stocks (2 × 10 7 cells/ml) were thawed and seeded into multiple 1 L spinner flasks containing 300 ml CD OptiCHO (Life Technologies; A11222) media supplemented with 8 mM L ‐glutamine (Corning; 25‐005‐CV).…”

Aflatoxin-specific Monoclonal Antibody Selection for Immunoaffinity Column Development

“…A suitable monoclonal cell line should be stable and productive in terms of both cell growth and antibody production to be able to sustain long-term production of the desired antibody. Media requirements of the cell line should be minimized without diminishing antibody production [10,25]. Most of the antibodies need to be purified prior to use.…”