2’14
CULTURE PROCESSES AND SCALE-UP
Thermocirculator controller controller
1. Attach thermocirculator with silicon tubing to reactor and reservoir, fill with water and set temperature to 3’1°C. 2. Connect medium recycle stericonnectors. 3. Place marprene tubing in the peristaltic pump head. 4. Add prewarmed medium to the reservoir vessel. 5. Leave for 4 h to equilibrate. 6. Takeasamplefromthereservoirandmeasurep using aninde~endently o manufacturer,^ instrucr set points to desired values.
spend inoculum (5.0 x IO9cells l-1 bed volume) in 800 m1 (bed void volume) medium. to inoculation vessel. 3. Connect to reactor vessel. 4. Gently swirl the inoculation vessel to ensure an even cell suspension. 5. Slowly pump the inoculum into the bed with air pressure (hand pump), being l not to introduce air bubbles into the system. and refill the bed twice. spension cells, start perfusing mediately at 40 m1 min-l or at a linear flow velocity of 2 cm m i d . or anchorage-dependent cells, leave stationary for2-4 h for the cells to attach before perfusing at 40 m1 min-l.
un the culture until the ma~imumproduct concentration is reached.