Biomass soft sensor for a Pichia pastoris fed‐batch process based on phase detection and hybrid modeling

A common control strategy for the production of recombinant proteins in Pichia pastoris using the alcohol oxidase 1 (AOX1) promotor is to separate the bioprocess into two main phases: biomass generation on glycerol and protein production via methanol induction. This study reports the establishment of a soft sensor for the prediction of biomass concentration that adapts automatically to these distinct phases. A hybrid approach combining mechanistic (carbon balance) and data‐driven modeling (multiple linear regression) is used for this purpose. The model parameters are dynamically adapted according to the current process phase using a multilevel phase detection algorithm. This algorithm is based on the online data of CO 2 in the off‐gas (absolute value and first derivative) and cumulative base feed. The evaluation of the model resulted in a mean relative prediction error of 5.52% and R ² of .96 for the entire process. The resulting model was implemented as a soft sensor for the online monitoring of the P. pastoris bioprocess. The soft sensor can be used for quality control and as input to process control systems, for example, for methanol control.