Scalable Production of Adenovirus Vectors - Adenovirus: Methods and Protocols - page 180

Fig. 3 Main components for bioreactor assembly. (Note: The base bottle and sampling system have steel caps
with two integrated tube connectors)

10. Verify all bioreactor assembly and connections after steriliza-
tion ( see Note 27 ).
11. Connect the bioreactor to the control unit as follows:
(a) Water inlet and outlet to the jacket.
(b) Cooler inlet and outlet to the control unit ( see Note 28 ).
(c) Temperature, pO 2 , and pH probes.
(d) Connect the motor.
(e) Connect the aeration system to the gas supply system and
start supplying N 2 to prepare pO 2 probe calibration.
(f) When temperature is below 50 °C, fill the jacket with
water and start temperature control ( see Note 29 ).
12. Replace MilliQ water with 80 % working volume of culture
medium ( see Note 30 ).
(a) Once temperature and pO 2 readings are stable, calibrate
the pO 2 probe ( see Note 31 ).
(b) Fill the base bottle with the base solution and connect to
the corresponding pump.
(c) Select the desired pO 2 and pH and switch on the unit
controllers ( see Note 32 ).
(d) When the desired pO 2 and pH are attained the bioreactor
is ready for cell inoculation.

The protocol to produce HAdV vectors is split into two points: (1)
bioreactor inoculation and cell growth and (2) cell infection and
virus harvesting. HEK 293 cells seed train can be prepared directly

3.3.2 HAdV Production