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Process Optimization using High Throughput Automated Micro-Bioreactors in Chinese Hamster Ovary Cell Cultivation

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Rational design and optimization of fed-batch and continuous fermentations.

Wenhui Zhang¹, Mehmet Inan, Michael M Meagher

¹Xoma Ltd., Berkey, CA.,USA.

Methods in Molecular Biology (Clifton, N.J.)

|October 24, 2007

Summary

This study offers a rational method for optimizing Pichia pastoris fermentations, detailing fed-batch and continuous processes for both Mut+ and Muts strains. It provides a practical guide for recombinant fermentation development and optimization.

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Area of Science:

Biotechnology
Microbial Fermentation
Process Optimization

Background:

Pichia pastoris is a widely used methylotrophic yeast for recombinant protein production.
Optimizing fermentation processes is crucial for maximizing yields and efficiency.
Both Mut+ and Muts Pichia pastoris strains have distinct methanol utilization characteristics impacting fermentation.

Purpose of the Study:

To provide rational design and optimization strategies for fed-batch and continuous Pichia pastoris fermentations.
To detail methods for glycerol batch, fed-batch, transition, and methanol-based continuous stirred tank reactor (CSTR) phases.
To establish a practical protocol for P. pastoris recombinant fermentation process development.

Main Methods:

Utilizing glycerol and methanol consumption models for process design.
Rationally designing cell density, broth volume, substrate feed rate, and phase duration.
Anchoring optimization by analyzing the impact of specific growth rate/dilution time on productivity.
Deriving equations for simulating processes with optimal parameters.

Main Results:

Detailed methods for optimizing fed-batch and continuous Pichia pastoris fermentation phases.
Integration of glycerol and methanol consumption models for rational process design.
Demonstration of how specific growth rate/dilution time impacts overall process productivity.

Conclusions:

The developed protocol offers a practical manual for P. pastoris recombinant fermentation process development.
The strategies are applicable for optimizing both Mut+ and Muts Pichia pastoris strains.
This approach serves as a valuable reference for the fermentation of other microorganisms.