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

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Feed development for fed-batch CHO production process by semisteady state analysis.

Sarwat F Khattak¹, Zizhuo Xing, Brian Kenty

¹Process Sciences Upstream, Technical Operations, Bristol-Myers Squibb Company, Syracuse, NY 13221, USA.

Biotechnology Progress

|December 17, 2009

Summary

This summary is machine-generated.

Maintaining Chinese hamster ovary (CHO) cell cultures in semisteady state reveals optimal nutrient levels for reduced waste. Constant volumetric feeding strategies also enhance process consistency over glucose-based methods.

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Area of Science:

Biotechnology
Cell Culture Engineering
Metabolic Engineering

Background:

Semisteady state cultures offer a controlled environment for studying cell physiology and optimizing media development.
Chinese hamster ovary (CHO) cells are critical for biopharmaceutical production, necessitating efficient culture methods.

Purpose of the Study:

To investigate the impact of nutrient concentrations in semisteady state CHO cell cultures on metabolic byproducts and growth.
To compare different feeding strategies, including fixed volumetric feeds versus glucose-based feeds, for fed-batch processes.

Main Methods:

Established two semisteady states in CHO cell cultures with varying glucose and glutamine concentrations (5 mM glucose/0.5 mM glutamine vs. 15 mM glucose/1.5 mM glutamine).
Compared semisteady state conditions with a standard fed-batch process using glucose-level-based feeding.
Evaluated alternative fixed volumetric feeding strategies (30% and 40% of initial volume).

Main Results:

Lower glucose and glutamine levels in semisteady state cultures significantly reduced lactate and ammonia production.
Higher nutrient concentrations in semisteady state cultures led to increased ammonia production (78%) and reduced cell growth rate (30%).
Fixed volumetric feeding strategies demonstrated greater robustness and consistency compared to traditional glucose-based feeding.

Conclusions:

Optimizing nutrient levels in semisteady state CHO cultures is key to minimizing metabolic waste.
Fixed volumetric feeding strategies represent a promising alternative for enhancing fed-batch process consistency and efficiency.
Semisteady state analysis provides valuable insights for feed enhancement and continuous improvement in bioprocessing.