stream of one seed reactor can be adapted to multiple induced

reactor systems, as shown for two induced reactors in Fig. 3

exemplarily.

6. For continuous adaptation phase, only lines marked in green

are opened at this level of cultivation (Fig. 3); all lines marked

in black (i.e., induction feed lines) are closed at this stage.

7. For continuous adaptation phase, only lines marked in green

are opened (Fig. 3); all lines marked in black (i.e., induction

feed lines) are closed at this stage.

3.2.3

Induction Phase

1. Once stage 1 is in equilibrium state (constant CO2 signals for

4 residence times at least), induction feed can be added.

2. Pumps need to be adjusted to desired volumetric rates.

3. All lines are opened at this stage (Fig. 4).

4. Efflux of stage 1 thus contains solely biomass, whereas efflux of

stage 2 contains cells including product (see Note 4).

Fig. 4 Process overview of the continuous cascaded cultivation. Reactor 1 is used for biomass formation.

Biomass streams are separated equivalent into reactors 2 and 3; Reactors 2 and 3 are used for recombinant

protein production. For screening of recombinant protein production, only process parameters in reactors

2 and 3 are varied; all lines (marked in green) are opened in this figure; Di ¼ dilution rate (1/h), Fi ¼ feed rate

(L/h), Xi ¼ biomass flow (L/h), Pi ¼ product flow (L/h), Vi ¼ volume (L)

232

Julian Kopp and Oliver Spadiut