Efficient feeding profile optimization for recombinant protein production using physiological information.

A multivariate study was performed aiming at the optimization of a recombinant rhamnose inducible E. coli induction system with alkaline phosphatase as target product. The effects of typical factors with impact on post- as well as pre-induction feeding rates were investigated with respect to the space-time yield of the target product. The goal was increased understanding as well as quantitative characterization of these factors with respect to their physiological impact on the model system. The optical density (OD) at which the culture was induced had a strong positive effect on the space-time yield. Pre-induction growth rate (k) had a second-order effect, while induction feed rate drop (J), a factor defining the linear post-induction feed rate, was interacting with (k). However, explanation of the observed effects to acquire more understanding regarding their effect on cell metabolism was not straight forward. Hence, the original process parameters were transformed into physiological more meaningful parameters and served as the basis for a multivariate data analysis. The observed variance with respect to observed volumetric activity was fully explained by the specific substrate uptake rate (q (s)) and induction OD, merging the process parameters pre-induction growth rate (k) and feed rate drop (J) into the physiological parameter specific substrate uptake rate (q (s)). After transformation of the response volumetric activity (U/ml) into the biomass specific activity (U/g(biomass)), the observed variance was fully explained solely by the specific substrate uptake rate (q (s)). Due to physiological multivariate data analysis, the interpretation of the results was facilitated and factors were reduced. On the basis of the obtained results, it was concluded that the physiological parameter q (s) rather than process parameters (k, J, induction OD) should be used for process optimization with respect to the feeding profile.