Development of a dual specific growth rate-based fed-batch process for production of recombinant human granulocyte...

A number of limitations exist for production of human granulocyte colony-stimulating factor (G-CSF) in Pichia pastoris. In this study, two different specific growth rates (0.015 h^-1, 0.01 h^-1) were used sequentially in the mixed substrate feeding period during methanol induction phase to enhance the G-CSF titer in the culture broth. Necessary parameters required for implementing the feeding strategy, such as specific product yield on biomass (Y_P/X) and maintenance coefficient (m) on glycerol, methanol, and sorbitol were estimated using continuous culture technique. Using this strategy, for the same volumetric productivity, about 20% increase in protein titer was achieved over that obtained from the run carried out at a single pre-set value of 0.015 h^-1 alone. Thus, implementation of higher specific growth rate (0.015 h^-1) set during initial stages of the methanol induction phase followed by a lower specific growth rate (0.01 h^-1) helped in achieving increased protein titers.