Single-use bioreactors are commonly used for seeding stainless steel bioreactors or for producing material. The profitability of this equipment has been well demonstrated for more than a decade, but few data on their scalability are published. In May 2010, Merck Serono Biodevelopment began a study to evaluate the performances of disposable bioreactors. Because different technologies were available, this study compared performances of several types of mixing. The evaluation was performed both for seeding application and for clinical material production. This study featured 20–50 L disposable bioreactors with gas flow-rate scaled-down from seeding and production bioreactors. A fed-batch process producing a highly glycosylated molecule was performed twice in four types of disposable bioreactors. The quality of the molecule together with molecule concentration and cell growth were compared among the three single-use technologies. These process performances were also compared to 250 L and 1,250 L bioreactors and to a 3.6 L glass development bioreactor. These comparisons allowed Merck Serono Biodevelopment to conclude on their uses and on the scalability (up and down) of these disposable systems. This study was completed by a characterization of liquid–liquid and gas–liquid transfers inside each disposable bioreactor to estimate their potential in terms of cell culture.
Cell culture was performed twice in each disposable bioreactor under the same conditions.
A vial of Chinese hamster ovary (CHO) cells was thawed in commercial medium in a static container. Cells were expanded every two or three days in agitated containers until reaching 8.6.109 cells.
Cell Growth in Disposable Bioreactors
A sterility test was performed at 37 °C in each disposable bag two days before inoculation. The cells were then inoculated in parallel in all disposable bioreactors at the minimum working volume provided by the supplier. Growth was then observed for two days. Working volume was increased to the maximum working volume at the same cell density between bioreactors. Cell growth at maximum working volume lasted three days.
Temperature was regulated at 37 °C. Stirring speed was fixed by the supplier.
Oxygen was regulated at 50% only with oxygen at a flow rate scaled-down from MSB 250 L seeding bioreactor for paddle and stirred mixing bioreactors and at a flow rate fixed by the supplier and MSB according to its experiences with the orbital shaking bioreactor.