ABSTRACT
Using one unified single-use cell culture platform for facilities that require culture of different types of cells can significantly
reduce equipment costs, the risk of cross-contamination, and the requirement for training, cleaning, sterilization, and validation.
Here, a bioreactor platform consisting of a presterilized, flexible, disposable plastic bag on a rocking platform and filled
with media and a gas mixture is shown to support the growth of E. coli, Chinese hamster ovary (CHO) cells, and hybridoma cells as measured by the daily cell density, nutrient consumption, and
production of metabolites.
Single-use disposable components offer many advantages in biologics manufacturing, and they collectively provide a platform
to culture diverse cell types in a facility. This is particularly important for multi-use contract manufacturing facilities
such as PacificGMP that service the needs of diverse clientele.
Disposable components include bioprocess bags, tubing, capsule filters, tangential flow capsules, bioreactors, chromatography
capsules, and mixing systems. They are supplied clean and ready to use, which obviates the need for sterilization and decreases
the requirement for services such as water for injection (WFI) water systems, and steam generators. Disposable components
are not used for subsequent operations, eliminating the chance of cross contamination between process runs. Because the need
for stainless-steel equipment is reduced or eliminated, long lead times for equipment installation can be avoided. Single-use
ystems are less complex, reducing engineering requirements. There is no need for clean-in-place (CIP) or steam-in-place (SIP)
operations, or any of the associated piping, valves, controls, or pressure rating of vessels. Moreover, the use of disposable
components reduces the complexity of validation. Because there are fewer reusable components, fewer items need to be tracked,
and extensive validation studies for sterilization and cleaning can be eliminated. Finally, by removing the limitations of
hard piping and stationary tanks, disposable components allow for operations to be more rapidly reconfigured for a new process
run.
There are several options for disposable, scalable bioprocess bags for completing upstream processing steps. The design of
a cell culture system will affect the shear forces experienced by cells as well as the mass transfer of gases, and different
types of cells will have different sensitivities to these parameters. Generally, mammalian cells are more sensitive to shear
forces than bacteria, but bacteria are more sensitive to oxygen limitation. The following studies demonstrate that the Wave
Bioreactor (GE Healthcare Chalfont St. Giles, UK) effectively supports the growth and productivity of different types of cells,
and the versatility of the platform is further demonstrated by the flexibility of feed strategies and scalability of the systems.
BIOREACTOR SET UP
Each bioreactor consists of a presterilized, flexible, disposable, plastic Cellbag that is placed on a rocking platform and
filled with up to half of the bag volume with media. The bioreactors used in these studies were the System20/50EH, System200,
and System1000 (GE Healthcare) with maximum culture volumes of 25, 100, and 500 L of media, respectively. The following naming
convention will be used to identify the culture set up: the volume of media in a Cellbag of a certain volume, e.g., 100/200
L indicates 100 L of media in the System200 bioreactor.
The remaining volume of the Cellbag is inflated with a process gas mixture, composed of carbon dioxide (CO2) and oxygen (O2), which is mixed with air in a controlled manner. These gases are added using a sterile inlet filter that is pre-attached
to the bag. Continuous airflow provides oxygenation and gas exchange for pH control and CO2 removal. Exhaust air passes through another sterile filter and a backpressure control valve. The backpressure control valve
ensures that the Cellbag is always fully inflated at any airflow and prevents overinflation.
Liquid mixing and mass transfer of gases are achieved by rocking the Cellbag back and forth at a user-determined rocking angle
and frequency. The waves generated by the rocking motion greatly increase surface area to enhance gas transfer. The wave motion
also promotes bulk mixing and off-bottom suspension of cells and particles without any damage to the cells. Temperature is
controlled by a heater located in the base plate of the unit that warms the underside of the bag. The heater is regulated
by a noninvasive temperature sensor that is also in the base plate of the unit.
