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More similar than different: Host cell protein production using three null CHO cell lines
Inn H Yuk1, Julie Nishihara2, Donald Walker2
1Early Stage Cell Culture, Genentech, 1 DNA Way, South San Francisco, California, 94080. inn@gene.com.
Biotechnology and Bioengineering
|April 21, 2015
Summary
Host cell protein (HCP) profiles in Chinese Hamster Ovary (CHO) cell culture harvests are surprisingly consistent across different cell lines and upstream processes. Even with declining cell viability, HCP levels remain stable, suggesting less diversity than previously assumed.
Area of Science:
- Biotechnology
- Biopharmaceutical Manufacturing
- Proteomics
Background:
- Host cell proteins (HCPs) are critical impurities in biopharmaceutical production.
- Understanding HCP diversity is essential for effective downstream processing and product safety.
- Chinese Hamster Ovary (CHO) cells are widely used for recombinant protein production.
Purpose of the Study:
- To investigate the diversity of HCP profiles in the cell culture harvest from different CHO cell lines.
- To compare HCP production dynamics and their correlation with cell viability.
- To evaluate analytical methods for HCP characterization.
Main Methods:
- Comparison of HCP profiles from three CHO cell lines using null runs.
- Time-course analysis of HCPs using multi-analyte ELISA.
- Correlation analysis with intracellular HCPs like phospholipase B-like 2 (PLBL2) and lactate dehydrogenase (LDH).
- Proteomic analysis using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS).
- Assessment of HCP profiles under declining culture viability.
Main Results:
- HCP profiles showed unexpected similarity across different CHO cell lines and upstream processes.
- Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) identified common HCPs across cell lines, despite differing 2D-PAGE patterns.
- Extracellular HCP levels remained relatively constant even with significant declines in cell viability.
- High-density CHO cultures can accumulate substantial amounts of immunogenic HCPs by harvest time.
Conclusions:
- CHO-derived HCPs in feedstock may exhibit less diversity across cell lines and processes than previously anticipated.
- HCP profiles are not dramatically altered by declining culture viability.
- LC-MS/MS offers advantages over ELISA and 2D-PAGE for comprehensive HCP analysis.
- Fed-batch CHO cultures can yield significant extracellular HCPs requiring efficient removal during downstream processing.
